EP1687018B1 - Hepatitis c virus inhibitors - Google Patents

Hepatitis c virus inhibitors Download PDF

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Publication number
EP1687018B1
EP1687018B1 EP04811049A EP04811049A EP1687018B1 EP 1687018 B1 EP1687018 B1 EP 1687018B1 EP 04811049 A EP04811049 A EP 04811049A EP 04811049 A EP04811049 A EP 04811049A EP 1687018 B1 EP1687018 B1 EP 1687018B1
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Prior art keywords
compound
alkyl
alkoxy
aryl
hcv
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English (en)
French (fr)
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EP1687018A4 (en
EP1687018A1 (en
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Paul Michael Scola
Fiona Mcphee
Nicholas A. Meanwell
Piyasena Hewawasam
Jeffrey Allen Campbell
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Bristol Myers Squibb Co
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Bristol Myers Squibb Co
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/08—Tripeptides
    • C07K5/0802—Tripeptides with the first amino acid being neutral
    • C07K5/0804—Tripeptides with the first amino acid being neutral and aliphatic
    • C07K5/0808—Tripeptides with the first amino acid being neutral and aliphatic the side chain containing 2 to 4 carbon atoms, e.g. Val, Ile, Leu
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A61P1/16—Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A61P31/14—Antivirals for RNA viruses
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/08—Tripeptides
    • C07K5/0802—Tripeptides with the first amino acid being neutral
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides

Definitions

  • the present invention is generally directed to antiviral compounds, and more specifically directed to compounds which inhibit the functioning of the NS3 protease (also referred to herein as "serine protease") encoded by Hepatitis C virus (HCV), compositions comprising such compounds and methods for inhibiting the functioning of the NS3 protease.
  • NS3 protease also referred to herein as "serine protease”
  • HCV Hepatitis C virus
  • HCV is a major human pathogen, infecting an estimated 170 million persons worldwide - roughly five times the number infected by human immunodeficiency virus type 1. A substantial fraction of these HCV infected individuals develop serious progressive liver disease, including cirrhosis and hepatocellular carcinoma. ( Lauer, G. M.; Walker, B. D. N. Engl. J. Med. (2001), 345, 41-52 ).
  • HCV is a positive-stranded RNA virus. Based on a comparison of the deduced amino acid sequence and the extensive similarity in the 5' untranslated region, HCV has been classified as a separate genus in the Flaviviridae family. All members of the Flaviviridae family have enveloped virions that contain a positive stranded RNA genome encoding all known virus-specific proteins via translation of a single, uninterrupted, open reading frame.
  • the single strand HCV RNA genome is approximately 9500 nucleotides in length and has a single open reading frame (ORF) encoding a single large polyprotein of about 3000 amino acids. In infected cells, this polyprotein is cleaved at multiple sites by cellular and viral proteases to produce the structural and non- structural (NS) proteins. In the case of HCV, the generation of mature non-structural proteins (NS2, NS3, NS4A, NS4B, NS5A, and NS5B) is effected by two viral proteases.
  • ORF open reading frame
  • the first one is believed to cleave at the NS2-NS3 junction; the second one is a serine protease contained within the N-terminal region of NS3 and mediates all the subsequent cleavages downstream of NS3, both in cis, at the NS3-NS4A cleavage site, and in trans, for the remaining NS4A- NS4B, NS4B-NS5A, NS5A-NS5B sites.
  • the NS4A protein appears to serve multiple functions, acting as a cofactor for the NS3 protease and possibly assisting in the membrane localization of NS3 and other viral replicase components.
  • NS3 protein The complex formation of the NS3 protein with NS4A seems necessary to the processing events, enhancing the proteolytic efficiency at all of the sites.
  • the NS3 protein also exhibits nucleoside triphosphatase and RNA helicase activities.
  • NS5B is a RNA-dependent RNA polymerase that is involved in the replication of HCV.
  • the present invention relates to compounds having the formula wherein:
  • the present invention also provides compositions comprising the compounds or pharmaceutically acceptable salts, or solvates thereof and a pharmaceutically acceptable carrier.
  • the present invention provides pharmaceutical compositions useful for inhibiting HCV NS3 comprising a therapeutically effective amount of a compound of the present invention, or a pharmaceutically acceptable salt, or solvate thereof, and a pharmaceutically acceptable carrier.
  • the present invention further provides methods for treating patients infected with HCV, comprising administering to the patient a therapeutically effective amount of a compound of the present invention, or a pharmaceutically acceptable salt, or solvate thereof. Additionally, the present invention provides methods of inhibiting HCV NS3 protease by contacting the NS3 protease with a compound of the present invention.
  • the present invention provides peptide compounds that can inhibit the functioning of the NS3 protease, e.g., in combination with the NS4A protease.
  • the present invention makes it possible to administer combination therapy to a patient whereby a compound in accordance with the present invention, which is effective to inhibit the HCV NS3 protease, can be administered with another compound having anti-HCV activity, e.g., a compound which is effective to inhibit the function of a target selected from the group consisting of HCV metalloprotease, HCV serine protease, HCV polymerase, HCV helicase, HCV NS4B protein, HCV entry, HCV assembly, HCV egress, HCV NS5A protein, IMPDH and a nucleoside analog for the treatment of an HCV infection.
  • a target selected from the group consisting of HCV metalloprotease, HCV serine protease, HCV polymerase, HCV helicase, HCV NS4B protein, HCV entry, HCV assembly, HCV egress, HCV NS5A protein, IMPDH and a nucleoside analog for the treatment of an HCV infection.
  • d and 1 or (+) and (-) are employed to designate the sign of rotation of plane-polarized light by the compound, with (-) or 1 meaning that the compound is levorotatory and (+) or d, meaning the compound; is dextrorotatory.
  • these compounds called stereoisomers, are identical except that they are mirror images of one another.
  • a specific stereoisomer of a mirror image pair may also be referred to as an enantiomer, and a mixture of such isomers is often called an enantiomeric mixture.
  • (R) or (S) it is to designate the absolute configuration of a substituent in context to the whole compound and not in context to the substituent alone.
  • racemic mixture and “racemate” refer to an equimolar mixture of two enantiomeric species, devoid of optical activity.
  • chiral refers to molecules which have the property of non-superimposability of the mirror image partner, while the term “achiral” refers to molecules which are superimposable on their mirror image partner.
  • stereoisomers refers to compounds which have identical chemical composition, but differ with regard to the arrangement of the atoms or groups in space.
  • diastereomer refers to a stereoisomer which is not an enantiomer, e.g., a stereoisomer with two or more centers of chirality and whose molecules are not mirror images of one another.
  • Diastereomers have different physical properties, e.g. melting points, boiling points, spectral properties, and reactivities. Mixtures of diastereomers may separate under high resolution analytical procedures such as electrophoresis and chromatography.
  • enantiomers refers to two stereoisomers of a compound which are non-superimposable mirror images of one another.
  • salts are intended to include nontoxic salts synthesized from a compound which contains a basic or acidic moiety by conventional chemical methods. Generally, such salts can be prepared by reacting the free acid or base forms of these compounds with a stoichiometric amount of the appropriate base or acid in water or in an organic solvent, or in a mixture of the two; generally, nonaqueous media like ether, ethyl acetate, ethanol, isopropanol, or acetonitrile are preferred. Lists of suitable salts are found in Remington's Pharmaceutical Sciences, 18th ed., Mack Publishing Company, Easton, PA, 1990, p. 1445 . The compounds of the present invention are useful in the form of the free base or acid or in the form of a pharmaceutically acceptable salt thereof. All forms are within the scope of the invention.
  • terapéuticaally effective amount means the total amount of each active component that is sufficient to show a meaningful patient benefit, e.g., a sustained reduction in viral load.
  • a meaningful patient benefit e.g., a sustained reduction in viral load.
  • the term refers to that ingredient alone.
  • the term refers to combined amounts of the active ingredients that result in the therapeutic effect, whether administered in combination, serially or simultaneously.
  • derivative means a chemically modified compound wherein the modification is considered routine by the ordinary skilled chemist, such as an ester or an amide of an acid, protecting groups, such as a benzyl group for an alcohol or thiol, and tert-butoxycarbonyl group for an amine.
  • solvate means a physical association of a compound of this invention with one or more solvent molecules, whether organic or inorganic. This physical association includes hydrogen bonding. In certain instances the solvate will be capable of isolation, for example when one or more solvent molecules are incorporated in the crystal lattice of the crystalline solid.
  • solvent encompasses both solution-phase and isolable solvates. Exemplary solvates include hydrates, ethanolates, methanolates, isopropanolates and the like.
  • a prodrug of a compound of the invention may be formed in a conventional manner with a functional group of the compounds such as with an amino, hydroxy or carboxy group when present.
  • the prodrug derivative form often offers advantages of solubility, tissue compatibility, or delayed release in a mammalian organism (see, Bundgard, H., Design of Prodrugs, pp. 7-9,21-24, Elsevier, Amsterdam 1985 ).
  • Prodrugs include acid derivatives well known to practitioners of the art, such as, for example, esters prepared by reaction of the parent acidic compound with a suitable alcohol, or amides prepared by reaction of the parent acid compound with a suitable amine.
  • patient includes both human and other mammals.
  • composition means a composition comprising a compound of the invention in combination with at least one additional pharmaceutical carrier, i. e., adjuvant, excipient or vehicle, such as diluents, preserving agents, fillers, flow regulating agents, disintegrating agents, wetting agents, emulsifying agents, suspending agents, sweetening agents, flavoring agents, perfuming agents, antibacterial agents, antifungal agents, lubricating agents and dispensing agents, depending on the nature of the mode of administration and dosage forms.
  • additional pharmaceutical carrier i. e., adjuents, preserving agents, fillers, flow regulating agents, disintegrating agents, wetting agents, emulsifying agents, suspending agents, sweetening agents, flavoring agents, perfuming agents, antibacterial agents, antifungal agents, lubricating agents and dispensing agents, depending on the nature of the mode of administration and dosage forms.
  • additional pharmaceutical carrier i. e., adjuvant, excipient or vehicle, such as diluents
  • phrases "pharmaceutically acceptable” is employed herein to refer to those compounds, materials, compositions, and/or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of patients without excessive toxicity, irritation, allergic response, or other problem or complication commensurate with a reasonable risk/benefit ratio.
  • treating refers to: (i) preventing a disease, disorder or condition from occurring in a patient which may be predisposed to the disease, disorder and/or condition but has not yet been diagnosed as having it; (ii) inhibiting the disease, disorder or condition, i.e., arresting its development; and (iii) relieving the disease, disorder or condition, i.e., causing regression of the disease, disorder and/or condition.
  • substituted includes substitution at from one to the maximum number of possible binding sites on the core, e.p., organic radical, to which the subsitutent is bonded, e.g., mono-, di-, tri- or tetra- substituted, unless otherwise specifically stated.
  • organic radicals e.g., hydrocarbons and substituted hydrocarbons
  • groups e.g., alkylalkoxyamine or arylalkyl
  • groups include all possible stable configurations, unless otherwise specifically stated. Certain radicals and combinations are defined below for purposes of illustration.
  • halo as used herein means a halogen substituent selected from bromo, chloro, fluoro or iodo.
  • haloalkyl means an alkyl group that in substituted with one or more halo substituents.
  • alkyl as used herein means acyclic, straight or branched chain alkyl substituents having the specified number of carbon atoms and includes, for example, methyl, ethyl, propyl, butyl, tert-butyl, hexyl, 1-methylethyl, 1-methylpropyl, 2-methypropyl, 1,1-dimethylethyl.
  • C 1-6 alkyl refers to an alkyl group having from one to six carbon atoms.
  • lower alkyl means an alkyl group having from one to six, preferably from one to four carbon atoms.
  • alkylester means an alkyl group additionally containing on ester group. Generally, a stated carbon number range, e.g., C 2-6 alkylester, includes all of the carbon atoms in the radical.
  • alkenyl as used herein means an alkyl radical containing at least one double bond, e.g., ethenyl (vinyl) and alkyl.
  • alkoxy as used herein means an alkyl group with the indicated number of carbon atoms attached to an oxygen atom. Alkoxy includes, for example, methoxy, ethoxy, propoxy, 1-methylethoxy, butoxy and 1,1-dimethylethoxy. The latter radical is referred to in the art as tert-butoxy.
  • alkoxycarbonyl means an alkoxy group additionally containing a carbonyl group.
  • cycloalkyl as used herein means a cycloalkyl substituent containing the indicated number of carbon atoms and includes, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl and spiro cyclic groups such as spirocyclopropyl as spirocyclobutyl.
  • cycloalkoxy as used herein means a cycloalkyl group linked to an oxygen atom, such as, for example, cyclobutyloxy or cyclopropyloxy.
  • alkylcycloalkyl means a cycloalkyl group linked to an alkyl group.
  • the stated carbon number range includes the total number of carbons in the radical, unless otherwise specfically stated.
  • This a C 4-10 alkylcycloalkyl may contain from 1-7 carbon atoms in the alkyl group and from 3-9 carbon atoms in the ring, e.g., cyclopropylmethyl or cyclohexylethyl.
  • aryl as used herein means an aromatic moiety containing the indicated number of carbon atoms, such as phenyl, indanyl or naphthyl.
  • C 6-10 aryl refers to an aromatic moiety having from six to ten carbon atoms which may be in the form of a monocyclic or bicyclic structure.
  • haloaryl refers to an aryl mono, di or tri substituted with one or more halogen atoms.
  • alkylaryl arylalkyl
  • aralalkyl mean an aryl group substituted with one or more alkyl groups. Unless the carbon range of each group is specified, the stated range applies to the entire substituent.
  • a C 7-14 alkylaryl group many have from 1-8 carbon atoms in the alkyl group for a monocyclic aromatic and from 1-4 carbon atoms in the alkyl group for a fused aromatic.
  • the attachment of the group to bonding site on the molecule can be either at the aryl group or the alkyl group.
  • aryl radicals include those substituted with typical substituents known to those skilled in the art, e.g., halogen, hydroxy, carboxy, carbonyl, nitro, sulfo, amino, cyano, dialkylamino haloalkyl, CF 3 , haloalkoxy, thioalkyl, alkanoyl, SH, alkylamino, alkylamide, dialkylamide, carboxyester, alkylsulfone, alkylsulfonamide and alkyl(alkoxy)amine.
  • alkylaryl groups include benzyl, butylphenyl and 1-naphthylmethyl.
  • alkanoyl as used herein means straight or branched 1-oxoalkyl radicals containing the indicated number of carbon atoms and includes, for example, formyl, acetyl, 1-oxopropyl (propionyl), 2-methyl-1-oxopropyl, 1-oxohexyl and the like.
  • alkylamide as used herein means an amide mono-substituted with an alkyl, such as
  • heterocycle also referred to as “Het”, as used herein means 7-12 membered bicyclic heterocycles and 5-7 membered monocyclic heterocycles.
  • Preferred bicyclic heterocycles are 7-12 membered fused bicyclic ring systems (both rings share an adjacent pair of atoms) containing from one to four heteroatoms selected from nitrogen, oxygen and sulfur, wherein both rings of the heterocycle are fully unsaturated.
  • the nitrogen and sulfur heteroatoms atoms may be optionally oxidized.
  • the bicyclic heterocycle may contain the heteroatoms in one or both rings. Unless a specific heterocycle is specified, e.g., a fluorinated 7-12 membered bicyclic heterocycle, or the heterocycle is stated to be unsubstituted, the heterocycles include those substituted with typical substituents known to those skilled in the art.
  • the bicyclic heterocycle may also contain substituents on any of the ring carbon atoms, e.g., one to three substituents.
  • suitable substituents include C 1-6 alkyl, C 3-7 cycloalkyl, C 1-6 alkoxy, C 3-7 cycloalkoxy, halo-C 1 - 6 alkyl, CF 3 , mono-or di- halo-C 1-6 alkoxy, cyano, halo, thioalkyl, hydroxy, alkanoyl, NO 2 , SH, , amino, C 1-6 alkylamino, di (C 1 - 6 ) alkylamino, di (C 1 - 6 ) alkylamide, carboxyl, (C 1-6 ) carboxyester, C 1-6 alkylsulfone, C 1-6 alkylsulfonamide, C 1-6 alkylsulfoxide, di (C 1-6 ) alkyl(alkoxy)amine, C 6-10
  • bicyclic heterocycle When two substituents are attached to vicinal carbon atoms of the bicyclic heterocycle, they can join to form a ring, e.g., a five, six or seven membered ring system containing up to two heteroatoms selecting from oxygen and nitrogen.
  • the bicyclic heterocycle may be attached to the molecule, e.g. R 1 in formula I, at any atom in the ring and preferably carbon.
  • bicyclic heterocycles include the following ring systems:
  • Preferred monocyclic heterocycles are 5-7 membered saturated, partially saturated or fully unsaturated ring system (this latter subset also herein referred to as unsaturated heteroaromatic) containing in the ring from one to four heteroatoms selected from nitrogen, oxygen and sulfur, wherein the sulfur and nitrogen heteroatoms may be optionally oxidized.
  • unsaturated heteroaromatic a specific heterocycle is specified, e.g., a C 1-6 alkoxy substituted 5-7 membered monocyclic heterocycle, or the heterocycle is stated to be unsubstituted
  • the heterocycles include those substituted with typical substituents known to those skilled in the art.
  • the monocyclic heterocycle may also contain substituents on any of the ring atoms, e.g., one to three substituents.
  • suitable substituents include C 1-6 alkyl, C 3-7 cycloalkyl, C 1-6 alkoxy, C 3-7 cycloalkoxy, halo-C 1-6 alkyl, CF 3 , mono-or di- halo-C 1-6 alkoxy, cyano, halo, thioalkyl, hydroxy, alkanoyl, NO 2 , SH, , amino, C 1-6 akylamino, di (C 1-6 ) alkylamino, di (C 1-6 ) alkylamide, carboxyl, (C 1-6 ) carboxyester, C 1-6 alkylsulfone, C 1-6 alkylsulfoxide, C 1-6 alkylsulfonamide, di (C 1-6 ) alkyl(alkoxy)amine, C 6-10 aryl, C 7
  • monocyclic heterocycles include the following (and their tautomers):
  • heterocycles used in the compounds of the present invention should be stable.
  • stable compounds are those which can be synthesized, isolated and formulated using techniques known the those skilled in the art without degradation of the compound.
  • substituted with reference to an amino acid or amino acid derivative means a radical derived from the corresponding ⁇ -amino acid.
  • substituents methyl, iso-propyl, and phenyl represent the amino acids alanine, valine, and phenyl glycine, respectively.
  • P1', P1, P2, P3 and P4" map the relative positions of the amino acid residues of a protease inhibitor binding relative to the binding of the natural peptide cleavage substrate.
  • Cleavage occurs in the natural substrate between P1 and P1' where the nonprime positions designate amino acids starting from the C-terminus end of the peptide natural cleavage site extending towards the N-terminus; whereas, the prime positions emanate from the N-terminus end of the cleavage site designation and extend towards the C-terminus.
  • P1' refers to the first position away from the right hand end of the C-terminus of the cleavage site (ie.
  • tert -butylglycine refers to a compound of the formula:
  • reduct with reference to an amino acid or amino acid derivative means a radical derived from the corresponding ⁇ -amino acid by eliminating the hydroxyl of the carboxy group and one hydrogen of the ⁇ -amino acid group.
  • the terms Gln, Ala, Gly, Ile, Arg, Asp, Phe, Ser, Leu, Cys, Asn, Sar and Tyr represent the "residues" of L -glutamine, L -alanine, glycine, L -isoleucine, L -arginine, L -aspartic acid, L -phenylalanine, L -serine, L -leucine, L -cysteine, L -asparagine, sarcosine and L -tyrosine, respectively.
  • side chain with reference to an amino acid or amino acid residue means a group attached to the ⁇ -carbon atom of the ⁇ -amino acid.
  • R-group side chain for glycine is hydrogen, for alanine it is methyl, for valine it is isopropyl.
  • specific R-groups or side chains of the ⁇ -amino acids reference is made to A.L. Lehninger's text on Biochemistry (see chapter 4).
  • R 1 and R 2 are each independently C 1-8 alkyl, C 3-7 cycloalkyl, C 4-10 alkylcycloalkyl, C 1-6 alkoxy, C 1-6 aryl, C 7-14 alkylaryl, 5-7 membered heteroaryl or C 7-14 alkylheteroaryl; or R 1 and R 2 , together with the nitrogen atom to which they are attached, join to form a 4-8 membered monocyclic heterocycle.
  • R 1 and R 2 are each independently C 1-8 alkyl, C 4-10 alkylcycloalkyl, C 1-6 alkoxy, C 6-10 aryl, C 7-14 alkylaryl, 5-7 membered heteroaryl or C 7-14 alkylheteroaryl, or R 1 and R 2 , together with the nitrogen atom to which they are attached, join to form a 5-6 membered monocyclic heterocycle.
  • R 3 is C 1-6 alkyl, C 2-6 alkenyl or C 3-7 cycloalkyl. More preferably, R 3 is C 2-6 alkenyl.
  • R 4 is C 1-8 alkyl optionally substituted with C 6 aryl, C 1-6 alkoxy, carboxy, hydroxy, aryloxy, C 7-14 alkylaryloxy, C 2-6 alkylester or C 8-15 alkylarylester; C 3-12 alkenyl; C 3-7 cycloalkyl; or C 4-10 alkylcycloalkyl. More preferably, R 4 is C 1-8 alkyl optionally substituted with C 1-6 alkoxy; or C 3-7 cycloalkyl.
  • Y is H.
  • R 5 is (i) C 1-10 alkyl optionally substituted with phenyl, carboxyl, C 1-6 alkanoyl, 1-3 halogen, hydroxy, C 1-6 alkoxy; (ii) C 3-7 cycloalkyl, C 3-7 cycloalkoxy, or C 4-10 alkylcycloalklyl; or (iii) C 6-10 aryl or C 7-16 arylalkyl, each optionally substituted with C 1-6 alkyl or halogen. More preferably, R 5 is (i) C 1-10 alkyle optionally substituted with 1-3 halogen or C 1-6 alkoxy; or (ii) C 3-7 cycloalkyl or C 4-10 alkylcycloalkyl.
  • R 6 is H or C 1-6 alkyl optionally substituted with 1-3 halogens. More preferably, R 6 is H.
  • X is O or NH.
  • R' is Het; or C 6-10 aryl optionally substituted with R a . More preferably, R' is Het. Even more preferably, the heterocycle contains 1 or 2 nitrogen atoms and optionally a sulfur atom or an oxygen atom in the ring. Preferably, the heterocycle is substituted with at least one of C 1-6 alkyl, C 1-6 alkoxy, halo, C 1-6 aryl, C 7-14 alkylaryl, or a 5-7 membered monocyclic heterocycle.
  • R a is C 1-6 alkyl, C 3-7 cycloalkyl, C 1-6 alkoxy, halo-C 1-6 alkyl, halo, amino, C 6 aryl, or a 5-7 membered monocyclic heterocycle.
  • R 1 and R 2 are each independently C 1-8 alkyl, C 4-10 alkylcycloalkyl, C 1-6 alkoxy, C 6-10 aryl, C 7-14 alkylaryl, 5-7 membered heteroaryl, C 7-14 alkylheteroaryl, or R 1 and R 2 , together with the nitrogen atom to which they are attached, join to form a 5-6 membered monocyclic heterocycle More preferably, R 1 and R 2 are each independently C 1-3 alkyl or C 1-3 alkoxy.
  • R' is a bicyclic heterocycle. More preferably, the heterocycle contains 1 or 2 nitrogen atoms and optionally a sulfur atom or an oxygen atom in the ring. Preferably, the heterocycle is substituted with at least one of C 1-6 alkyl, C 1-6 alkoxy, halo, C 6 aryl, and a 5-7 membered monocyclic heterocycle. Even more preferably, R' is a bicyclic heterocycle containing 1 nitrogen atom and substituted with methoxy and at least one of a C 6 aryl and a 5-7 membered monocyclic heterocycle.
  • R' is a monocyclic heterocycle.
  • the heterocycle contains 1 or 2 nitrogen atoms and optionally a sulfur atom or an oxygen atom in the ring. More preferably, the heterocycle is substituted with at least one of C 1-6 alkyl, C 1-6 alkoxy, halo, C 6-10 aryl, C 7-14 alkylaryl, or a 5-7 membered monocyclic heterocycle.
  • R' is a monoyclic heterocycle containing 1 or 2 nitrogen atoms and substituted with methoxy and at least one of a C 6 aryl and a 5-7 membered monocyclic heterocycle.
  • R 1 and R 2 are each independently C 1-3 alkyl, or C 1-3 alkoxy. More preferably, R 1 is methyl. Preferably, R 2 is methyl or methoxy. Preferably, R 3 is vinyl. Preferably, R 4 is t-butyl. Preferably, R 5 is t-butyl. Preferably, R' is quinoline or isoquinoline optionally substituted with R 8 .
  • R 1 is methyl
  • R 2 is methoxy
  • R 3 is vinyl
  • R 4 is t-butyl
  • R 5 is t-butyl
  • R' is isoquinoline substituted with at least one R a .
  • R a is C 1-6 alkoxy. More preferably, R a further includes at least one of C 6 aryl or a 5-7 membered monocyclic heterocycle.
  • the compounds of the present invention which contain a basic moiety, can form salts by the addition of a pharmaceutically acceptable acid.
  • the acid addition salts are formed from a compound of Formula I and a pharmaceutically acceptable inorganic acid, including hydrochloric, hydrobromic, hydroiodic, sulfuric, phosphoric, or organic acid such as p-toluenesulfonic, methanesulfonic, acetic, benzoic, citric, malonic, fumaric, maleic, oxalic, succinic, sulfamic, or tartaric.
  • examples of such pharmaceutically acceptable salts include chloride, bromide, iodide, sulfate, phosphate, methanesulfonate, citrate, acetate, malonate, fumarate, sulfamate, and tartrate.
  • Salts of an amine group may also comprise quaternary ammonium salts in which the amino nitrogen carries a suitable organic group such as an alkyl, alkenyl, alkynyl or aralkyl moiety.
  • Base addition salts include those derived from inorganic bases which include, for example, alkali metal salts (e.g. sodium and potassium), alkaline earth metal salts (e.g. calcium and magnesium), aluminum salts and ammonium salts.
  • alkali metal salts e.g. sodium and potassium
  • alkaline earth metal salts e.g. calcium and magnesium
  • aluminum salts e.g. aluminum salts and ammonium salts.
  • suitable base addition salts include salts of physiologically acceptable organic bases such as trimethylamine, triethylamine, morpholine, pyridine, piperidine, picoline, dicyclohexylamine, N,N'-dibenzylethylenediamine, 2-hydroxyethylamine, bis-(2-hydroxyethyl)amine, tri-(2-hydroxyethyl)amine, procaine, dibenzylpiperidine, N-benzyl-(3-phenethylarnine, dehydroabietylamine, N,N'-bishydroabietylamine, glucamine, N-methylglucamine, collidine, quinine, quinoline, ethylenediamine, ornithine, choline, N,N'-benzylphenethylamine, chloroprocaine, diethanolamine, diethylamine, piperazine, tris(hydroxymethyl)aminomethane and tetramethylammonium hydroxide and basic
  • Certain compounds of the present invention, and their salts may also exist in the form of solvates with water, for example hydrates, or with organic solvents such as methanol, ethanol or acetonitrile to form, respectively, a methanolate, ethanolate or acetonitrilate.
  • the present invention includes each solvate and mixtures thereof.
  • compounds of the present invention may exhibit polymorphism.
  • the present invention also encompasses any such polymorphic form.
  • the compounds of the present invention also contain two or more chiral centers.
  • the compounds may include P1 cyclopropyl element of formula wherein C 1 and C 2 each represent an asymmetric carbon atom at positions 1 and 2 of the cyclopropyl ring.
  • the presence of these two asymmetric centers means that the compounds can exist as racemic mixtures of diastereomers, such as the diastereomers wherein R 2 is configured either syn to the amide or syn to the carbonyl as shown below.
  • the present invention includes both enantiomers and mixtures of enantiomers such as racemic mixtures.
  • the enantiomers may be resolved by methods known to those skilled in the art, for example, by formation of diastereoisomeric salts which may be separated by crystallization, gas-liquid or liquid chromatography, selective reaction of one enantiomer with an enantiomer-specific reagent. It will be appreciated that where the desired enantiomer is converted into another chemical entity by a separation technique, then an additional step is required to form the desired enantiomeric form. Alternatively, specific enantiomers may be synthesized by asymmetric synthesis using optically active reagents, substrates, catalysts or solvents, or by converting one enantiomer into the other by asymmetric transformation.
  • Certain compounds of the present invention may also exist in different stable conformational forms which may be separable. Torsional asymmetry due to restricted rotation about an asymmetric single bond, for example because of steric hindrance or ring strain, may permit separation of different conformers.
  • the present invention includes each conformational isomer of these compounds and mixtures thereof.
  • Certain compounds of the present invention may exist in zwitterionic form and the present invention includes each zwitterionic form of these compounds and mixtures thereof.
  • the starting materials useful to synthesize the compounds of the present invention are known to those skilled in the art and can be readily manufactured or are commercially available.
  • the compounds of the present invention can be manufactured by methods known to those skilled in the art, see e.g., US Patent No. 6,323,180 and US Patent Application Publication No. 20020111313 A1, published August 15, 2002 .
  • the following methods set forth below are provided for illustrative purposes. It will be recognized that it may be preferred or necessary to prepare such a compound in which a functional group is protected using a conventional protecting group then to remove the protecting group to provide a compound of the present invention.
  • the details concerning the use of protecting groups in accordance with the present invention are known to those skilled in the art.
  • the compounds of the present invention may, for example, be synthesized according to a general process as illustrated in Scheme I (wherein CPG is a carboxyl protecting group and APG is an amino protecting group):
  • the P1, P2, and P3 can be linked by well known peptide coupling techniques.
  • the P1, P2, and P3 groups may be linked together in any order as long as the final compound corresponds to peptides of the invention.
  • P3 can be linked to P2-P1; or P1 linked to P3-P2.
  • peptides are elongated by deprotecting the ⁇ -amino group of the N-terminal residue and coupling the unprotected carboxyl group of the next suitably N-protected amino acid through a peptide linkage using the methods described. This deprotection and coupling procedure is repeated until the desired sequence is obtained. This coupling can be performed with the constituent amino acids in stepwise fashion, as depicted in Scheme I.
  • Coupling between two amino acids, an amino acid and a peptide, or two peptide fragments can be carried out using standard coupling procedures such as the azide method, mixed carbonic-carboxylic acid anhydride (isobutyl chloroformate) method, carbodiimide (dicyclohexylcarbodiimide, diisopropylcarbodiimide, or water-soluble carbodiimide) method, active ester (p-nitrophenyl ester, N-hydroxysuccinic imido ester) method, Woodward reagent K-method, carbonyldiimidazole method, phosphorus reagents or oxidation-reduction methods. Some of these methods (especially the carbodiimide method) can be enhanced by adding 1-hydroxybenzotriazole or 4-DMAP. These coupling reactions can be performed in either solution (liquid phase) or solid phase.
  • the coupling step involves the dehydrative coupling of a free carboxyl of one reactant with the free amino group of the other reactant in the present of a coupling agent to form a linking amide bond.
  • coupling agents are found in general textbooks on peptide chemistry, for example, M. Bodanszky, "Peptide Chemistry", 2nd rev ed, Springer-Verlag, Berlin, Germany, (1993 ).
  • suitable coupling agents are N,N'-dicyclohexylcarbodiimide, 1-hydroxybenzotriazole in the presence of N,N'-dicyclohexylcarbodiimide or N-ethyl-N'-[(3-dimethylamino)propyl]carbodiimide.
  • a practical and useful coupling agent is the commercially available (benzotriazol-1-yloxy)tris-(dimethylamino)phosphonium hexafluorophosphate, either by itself or in the present of 1-hydroxybenzotriazole or 4-DMAP.
  • Another practical and useful coupling agent is commercially available 2-(1H-benzotriazol-1-yl)-N, N, N', N'-tetramethyluronium tetrafluoroborate. Still another practical and useful coupling agent is commercially available O-(7-azabenzotrizol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate.
  • the coupling reaction is conducted in an inert solvent, e.g. dichloromethane, acetonitrile or dimethylformamide.
  • An excess of a tertiary amine e.g.
  • reaction temperature usually ranges between 0 °C and 50 °C and the reaction time usually ranges between 15 min and 24 h.
  • APG The ⁇ -amino group of each amino acid to be coupled to the growing peptide chain must be protected (APG). Any protecting group known in the art can be used. Examples of such groups include: 1) acyl groups such as formyl, trifluoroacetyl, phthalyl, and p-toluenesulfonyl; 2) aromatic carbamate groups such as benzyloxycarbonyl (Cbz or Z) and substituted bensyloxycarbonyls, and 9-fluorenylmethyloxycarbonyl (Fmoc); 3) aliphatic carbamate groups such as tert-butyloxycarbonyl (Boc), ethoxycarbonyl, diisopropylmethoxycarbonyl, and allyloxycarbonyl; 4) cyclic alkyl carbamate groups such as cyclopentyloxycarbonyl and adamantyloxycarbonyl; 5) alkyl groups such as triphenylmethyl and
  • the preferred ⁇ -amino protecting group is either Boc or Fmoc. Many amino acid derivatives suitably protected for peptide synthesis are commercially available. The ⁇ -amino protecting group of the newly added amino acid residue is cleaved prior to the coupling of the next amino acid.
  • Boc group the methods of choice are trifluoroacetic acid, neat or in dichloromethane, or HCl in dioxane or in ethyl acetate.
  • the resulting ammonium salt is then neutralized either prior to the coupling or in situ with basic solutions such as aqueous buffers, or tertiary amines in dichloromethane or acetonitrile or dimethylformamide.
  • the reagents of choice are piperidine or substituted piperidine in dimethylformamide, but any secondary amine can be used.
  • the deprotection is carried out at a temperature between 0°C and room temperature (rt or RT) usually 20-22°C.
  • any of the amino acids having side chain functionalities must be protected during the preparation of the peptide using any of the above-described groups.
  • Those skilled in the art will appreciate that the selection and use of appropriate protecting groups for these side chain functionalities depend upon the amino acid and presence of other protecting groups in the peptide. The selection of such protecting groups is important in that the group must not be removed during the deprotection and coupling of the ⁇ -amino group.
  • Boc when used as the ⁇ -amino protecting group, the following side chain protecting group are suitable: p -toluenesulfonyl (tosyl) moieties can be used to protect the amino side chain of amino acids such as Lys and Arg; acetamidomethyl, benzyl (Bn), or tert -butylsulfonyl moieties can be used to protect the sulfide containing side chain of cysteine; bencyl (Bn) ethers can be used to protect the hydroxy containing side chains of serine, threonine or hydroxyproline; and benzyl esters can be used to protect the carboxy containing side chains of aspartic acid and glutamic acid.
  • p -toluenesulfonyl (tosyl) moieties can be used to protect the amino side chain of amino acids such as Lys and Arg
  • Fmoc is chosen for the ⁇ -amine protection
  • usually tert -butyl based protecting groups are acceptable.
  • Boc can be used for lysine and arginine, tert-butyl ether for serine, threonine and hydroxyproline, and tert -butyl ester for aspartic acid and glutamic acid.
  • Triphenylmethyl (Trityl) moiety can be used to protect the sulfide containing side chain of cysteine.
  • Cyclopentanol is treated with phosgene to furnish the corresponding chloroformate.
  • the chloroformate is treated with the desired NH 2 -tripeptide in the presence of a base such as triethylamine to afford the cyclopentylcarbamate.
  • the ⁇ -carboxyl group of the C-terminal residue is usually protected as an ester (CPG) that can be cleaved to give the carboxylic acid.
  • CPG ester
  • Protecting groups that can be used include: 1) alkyl esters such as methyl, trimethylsilylethyl and t-butyl, 2) aralkyl esters such as benzyl and substituted benzyl, or 3) esters that can be cleaved by mild base treatment or mild reductive means such as trichloroethyl and phenacyl esters.
  • the resulting ⁇ -carboxylic acid (resulting from cleavage by mild acid, mild base treatment or mild reductive means) is coupled with a R 1 (R 2 )NSO 2 NH 2 [prepared by described herein] in the presence of peptide-coupling agent such as CDI in the presence of a base such as LiHMDS (lithium bis-hexamethyldisilylamide) to incorporate the P1' moiety, effectively assembling the tripeptide P1'-P1-P2-P3-APG.
  • peptide-coupling agent such as CDI
  • a base such as LiHMDS (lithium bis-hexamethyldisilylamide)
  • Scheme II further shows the general process wherein compounds of Formula I are constructed by the coupling of tripeptide carboxylic acid intermediate (1) with a P1' sulfamide.
  • Said coupling reaction requires treatment of carboxylic acid (1) with a coupling reagent such as carbonyl diimidazole in a solvent such as THF, which can be heated to reflux, followed by the addition of the formed derivative of (1), to the P1' sulfamide, in a solvent such as THF or methylene chloride in the presence of a base such as lithium bishexamethyldisilylamide.
  • TFA amine salt can be directly employed in the subsequent coupling reaction or as an alternative the TFA amine salt can be first converted to the HCl amine salt, and this HCl amine salt is used in said coupling reaction as shown in Scheme III.
  • the coupling of said HCl amine salt (3) with the carboxyl terminus a P4-P3-P2 intermediate can be achieved using coupling reagents, such as HATU, in solvents such as dichloromethane to provide compounds of Formula I (4).
  • the resulting trifluoroacetic acid salt can be coupled with the carboxyl terminus of the P4-P3 element using standard coupling agents such as PyBop in the presence of base such as diisopropyl amine, and using solvents such methylene chloride to provide compounds of Formula I (4).
  • the P4-P3-P2 intermediate utilized in the above schemes can be constructed as previously described with a further description of this process shown in general Scheme V.
  • the free carboxyl terminus of the P4-P3 intermediate (1) can be coupled to the amino terminus of the P2 element to provide the P4-P3-P2 dipeptide (2).
  • the carboxyl terminus of the P4-P3-P2 intermediate can be deprotected by saponification of the ester group to provide P4-P3-P2 as the free carboxylic acid (3).
  • Intermediates like (3) can be converted to compounds of Formula I using the methods described herein.
  • intermediate (2) can be used as starting materials for the preparation of compounds of Formula I wherein the P3 group is capped with an amide or a sulfonamide, or thiourea, or a sulfamide.
  • the construction of said compounds of Formula I can be achieved using standard conditions for the formation of said P4 functionalities from amines.3
  • P1' terminus is incorporated into the molecules using one of the general processes outlined above and described in more detail below.
  • These P1' sulfamides can be prepared in a two step process using chlororsulfonylisocyanate as starting material. Said isocynate can be hydrolyzed to the corresponding chlorosulfamoyl chloride by treatment with water in a solvent such as THF. The intermediate sulfamoylchloride upon treatment with an amine, in the presence of a base, provides the desired sulfamide derivatives.
  • the P1 elements utilized in generating compounds of Formula I are in some cases commercially available, but are otherwise synthesized using the methods described herein and subsequently incorporated into compounds of Formula I using the methods described herein.
  • the substituted P1 cyclopropylamino acids can be synthesized following the general process outline in Scheme VIII.
  • this reaction is selective in that one of the enantiomers undergoes the reaction at a much greater rate than its mirror image providing for a kinetic resolution of the intermediate racemate.
  • the more preferred stereoisomer for integration into compounds of Formula I is 5a which houses the (1R, 2S) stereochemistry.
  • this enantiomer does not undergo ester cleavage and thereby this enantiomer 5a is recovered from the reaction mixture.
  • the less preferred enantiomer ,5b with houses the (1S, 2R) stereochemistry undergoes ester cleavage, i.e., hydrolysis, to provide the free acid 6.
  • the ester 5a can be separated from the acid product 6 by routine methods such as, for example, aqueous extraction methods or chromotography.
  • Scheme IX shows the coupling of an N-protected C4-hydroxyproline moiety with a heterocycle to form intermediate (4) and the subsequent modification of said intermediate (4) to a compound of Formula I by the process of peptide elongation as described herein.
  • a base is employed in the first step.
  • this coupling can be done using bases such as potassium tert-butoxide, or sodium hydride, in solvent such as DMF or DMSO or THF.
  • This coupling to the isoquinoline ring system occurs at the C1 positions (numbering for isoquinoline ring system shown in intermediate 2 of Scheme IX) and is directed by the chloro group which is displaced in this process.
  • the alternative leaving groups can be utilized at this position such as a fluoro as shown in the Scheme.
  • Said fluoro intermediates (3) are available from the corresponding chloro compound using literature procedures described herein.
  • the position of the leaving group (chloro or fluoro) in a given ring system can vary as shown in Scheme X, wherein the leaving group (fluoro in this example) is in the C6 position of the isoquinoline ring system of intermediate (2).
  • isoquinolines are incorporated into the final compounds and specifically into the P2 region of said compounds.
  • One skilled in the art would recognize that a number of general methods are available for the synthesis of isoquinolines. Moreoever, said isoquinolines generated by these methods can be readily incorporated into final compounds of Formula I using the processes described herein.
  • One general methodology for the synthesis of isoquinolines is shown in Scheme XII, wherein cinnamic acid derivatives, shown in general form as structure (2) are converted to 1-chloroisoquinolines in a four step process. Said chloroisoquinolines can be subsequently used in coupling reactions to C4-hydroxyproline derivatives as described herein.
  • acylazide (3) is coverted to the corresponding isoquinolone (4) as shown in the Scheme.
  • acylazide is heated to a temperature of approximately 190 degress celcius in a high boiling solvent such a diphenylmethane.
  • This reaction is general and provides moderate to good yields of substituted isoquinolone from the corresponding cinnamic acid derivatives.
  • said cinnamic acid derivatives are available commercially or can be obtained from the corresponding benzaldehyde (1) derivative by direct condensation with malonic acid or derivatives thereof and also by employing a Wittig reaction.
  • the intermediate isoquinolones (4) of Scheme XII can be converted to the corresponding 1-chloroisoquinoline by treatment with phosphorous oxychloride.
  • This reaction is general and can be applied to any of the isoquinolones, quinolones or additional heterocycles as shown herein to covert a hydroxy substituent to the corresponding chloro compound when said hydroxy is in conjugation with a nitrogen atom in said heterocylic ring systems.
  • the intermediate isoquinolines (3) can be converted to the corresponding 1-chloroquinolines (5) in a two step process as shown.
  • the first step in this sequence is the formation of the isoquinoline N-oxide(4) by treatment of isoquinoline (3) with meta-chloroperbenzoic acid in an aprotic solvent such as dichloromethane.
  • Intermediate (4) can be converted to the corresponding 1-chloroquinoline by treatment with phosphorous oxychloroide in refluxing chloroform.
  • this two step process is general and can be employed to procure chloroisoquinolines and chloroquinolines from the corresponding isoquinolines and quinolines respectively.
  • Scheme XIV Another method for the synthesis of the isoquinoline ring system is shown in Scheme XIV.
  • an ortho-alkylbenzamide derivative (1) is treated with a strong base such as tert-butyl lithium in a solvent such as THF at low temperature.
  • a nitrile derivative which undergoes an addition reaction with the anion derived from deprotonation of (1), resulting in the formation of (2).
  • This reaction is general and can be used for the formation of substituted isoquinolines.
  • Intermediate (2) of Scheme XIV can be converted to the corresponding 1-chloroquinoline by the methods described herein.
  • An additional method for the synthesis of isoquinolines is shown in Scheme XV.
  • intermediate (1) using tert-butyl lithium is described above.
  • said intermediate anion is trapped by an ester, resulting in the formation of intermediate (2) as shown below.
  • ketone (2) is condensed with ammoniumn acetate at elevated temperature providing for the formation of quinolone (3).
  • This reaction is general and can be applied for the construction of substituted isoquinolones which can then be converted to the corresponding 1-chloroisoquinolines as described herein.
  • Scheme XVII shows the conversion of a 1-chloro- 6-fluoro-isoquinoline to the corresponding 1-chloro-6-alkoxy-isoquinoline species, by treatment of (1) of (eq. 1) with a sodium or potassium alkoxide species in the alcohol solvent from which the alkoxide is derived at room temperature. In some cases it may be necessary to heat the reaction to drive it to completion.
  • Said chloroquinoline can be incorporated into a compound of Formula I using the art described herein.
  • Modifications of a P2 heterocyclic element can also be done after it's incorporation into compounds of Formula I as shown in (eq.2) of Scheme VXII.
  • compounds such as (1) in (eq. 2) which contain a leaving group in the pthalazine nucleus can be displaced by a nucleophile such as an alkoxide in solvents such as the corresponding alcohol from which the alkoxide is derived.
  • a nucleophile such as an alkoxide in solvents such as the corresponding alcohol from which the alkoxide is derived.
  • Scheme XVIII provides a general example for the modification of heterocycles as defined herein by employing palladium mediated coupling reactions.
  • Said couplings can be employed to functionalize a heterocycle at each position of the ring system providing said ring is suitably activated or functionalized, as for example with a chloride as shown in the Scheme.
  • This sequence begins with 1-chloroisoquinoline (1) which upon treatment with metachloroperbenzoic acid can be converted to the corresponding N-oxide (2).
  • Said intermediate (2) can be converted to the corresponding 1,3-dichloroisoquinoline (3) by treatment with phosphorous oxychloride in refluxing chloroform.
  • Intermediate (3) can be coupled with N-Boc-4-hydroxyproline by the methods described herein to provide intermediate (5) as shown in the Scheme.
  • Intermediate (5) can undergo a Suzuki coupling with an aryl boronic acid, in the presence of a palladium reagent and base, and in a solvent such as THF or toluene or DMF to provide the C3-arylisoquinoline intermediate (6).
  • Heteroarylboronic acids can also be employed in this Pd mediated coupling process to provide C3-heteroarylisoquinolines.
  • Intermediate (6) can be converted into final compounds of Formula I by the methods described herein.
  • the isoquinoline ring system of intermediate (3) can be further functionalized by employing either Suzuki couplings (Process 1: subjecting (3) to heteroaryl or aryl boronic acids in the presence of a palladium catalyst such as palladium tetra(triphenylphosphine) and a base such as cesium carbonate in solvents such as DMF) or Stille couplings (Process 2: subjecting (3) to heteroaryl or aryl tin dervatives in the presence of palladium catalyst such as palladium tetra(triphenylphosphine in solvents such as toluene).
  • Palladium reactions can also be employed to couple C4-amino proline elements with functionalized heterocycles.
  • Scheme XX shows intermediate (1) coupling with a functionalized isoquinoline in the presence of a palladium catalyst and a base in a solvent such as toluene.
  • Intermediates like (3) can be converted to compounds of Formula I using the methods described herein.
  • WO 03/099274 provides extensive examples and teachings on the prepration of proline derivatives as intermediates and these intermediates can be used in the preparation of compound of Formula 1.
  • compositions comprising a compound of the present invention, or a pharmaceutically acceptable salt, solvate or prodrug thereof, and a pharmaceutically acceptable carrier.
  • Pharmaceutical compositions of the present invention comprise a therapeutically effective amount of a compound of the invention, or a pharmaceutically acceptable salt, or solvate thereof, and a pharmaceutically acceptable carrier, with a pharmaceutically acceptable carrier, e.g., excipient, or vehicle diluent.
  • the active ingredient, i.e., compound, in such compositions typically comprises from 0.1 weight percent to 99.9 percent by weight of the composition, and often comprises from about 5 to 95 weight percent.
  • a composition comprising the compound of formula 1 and a pharmaceutically acceptable carrier.
  • the composition further comprises a compound having anti-HCV activity.
  • anti-HCV activity means the compound is effective to inhibit the function of a target selected from the group consisting of HCV metalloprotease, HCV serine protease, HCV polymerase, HCV helicase, HCV NS4B protein, HCV entry, HCV assembly, HCV egress, HCV NS5A protein, IMPDH and a nucleoside analog for the treatment of an HCV infection.
  • the other compound having anti-HCV activity is effective to inhibit the function of target in the HCV life cycle other than the HCV serine protease.
  • the compound having anti-HCV activity is an interferon.
  • the interferon is selected from the group consisting of interferon alpha 2B, pegylated interferon alpha, consensus interferon, interferon alpha 2A and lymphoblastiod interferon tau.
  • the compound having anti-HCV activity is selected from the group consisting of interleukin 2, interleukin 6, interleukin 12, a compound that enhances the development of a type 1 helper T cell response, interfering RNA, anti-sense RNA, Imiqimod, ribavirin, an inosine 5'-monophospate dehydrogenase inhibitor, amantadine, and rimantadine.
  • the composition comprises a compound of the invention, an interferon and ribavirin.
  • the compound having anti-HCV activity is a small molecule compound.
  • small molecule compound means a compound having a molecular weight of less than 1,500 daltons, preferably less than 1000 daltons.
  • the small molecule compound is effective to inhibit the function of a target selected from the group consisting of HCV metalloprotease, HCV serine protease, HCV polymerase, HCV helicase, HCV NS4B protein, HCV entry, HCV assembly, HCV egress, HCV NS5A protein, inosine monophophate dehydrogenase ("IMPDH”) and a nucleoside analog for the treatment of an HCV infection.
  • IMPDH inosine monophophate dehydrogenase
  • Table 1 below lists some illustrative examples of compounds that can be administered with the compounds of this invention.
  • the compounds of the invention can be administered with other anti-HCV activity compounds in combination therapy, either jointly or separately, or by combining the compounds into a composition.
  • compositions of this invention may be administered orally, parenterally or via an implanted reservoir. Oral administration or administration by injection are preferred. In some cases, the pH of the formulation may be adjusted with pharmaceutically acceptable acids, bases or buffers to enhance the stability of the formulated compound or its delivery form.
  • parenteral as used herein includes subcutaneous, intracutaneous, intravenous, intramuscular, intra-articular, intrasynovial, intrasternal, intrathecal, and intralesional injection or infusion techniques.
  • the pharmaceutical compositions of this invention may be administered in any orally acceptable dosage form including, capsules, tablets, and aqueous suspensions and solutions.
  • carriers which are commonly used include lactose and corn starch.
  • Lubricating agents, such as magnesium stearate, are also typically added.
  • useful diluents include lactose and dried corn starch.
  • aqueous suspensions are administered orally, the active ingredient is combined with emulsifying and suspending agents. If desired, certain sweetening and/or flavoring and/or coloring agents may be added.
  • suitable carriers for the above noted compositions can be found in standard pharmaceutical texts, e.g. in " Remington's Pharmaceutical Sciences", 19th ed., Mack Publishing Company, Easton, Penn., 1995 .
  • compositions of this invention can be prepared by known procedures using well-known and readily available ingredients.
  • the compositions of this invention may be formulated so as to provide quick, sustained or delayed release of the active ingredient after administration to the patient by employing procedures well known in the art.
  • the active ingredient will usually be admixed with a carrier, or diluted by a carrier, or enclosed within a carrier which may be in the form of a capsule, sachet, paper or other container.
  • the carrier serves as a diluent, it may be a solid, semi-solid or liquid material which acts as a vehicle, excipient or medium for the active ingredient.
  • compositions can be in the form of tablets, pills, powders, beadlets, lozenges, sachets, elixirs, suspensions, emulsions, solutions, syrups, aerosols, (as a solid or in a liquid medium), soft and hard gelatin capsules, suppositories, sterile injectable solutions, sterile packaged powders and the like. Further details concerning the design and preparation of suitable delivery forms of the pharmaceutical compositions of the invention are known to those skilled in the art.
  • Dosage levels of between about 0.01 and about 1000 milligram per kilogram (“mg/kg”) body weight per day, preferably between about 0.5 and about 250 mg/kg body weight per day of the compounds of the invention are typical in a monotherapy for the prevention and treatment of HCV mediated disease.
  • the pharmaceutical compositions of this invention will be administered from about 1 to about 5 times per day or alternatively, as a continuous infusion. Such administration can be used as a chronic or acute therapy.
  • the amount of active ingredient that may be combined with the carrier materials to produce a single dosage form will vary depending upon the host treated and the particular mode of administration.
  • compositions of this invention comprise a combination of a compound of the invention and one or more additional therapeutic or prophylactic agent
  • both the compound and the additional agent are usually present at dosage levels of between about 10 to 100%, and more preferably between about 10 and 80% of the dosage normally administered in a monotherapy regimen.
  • composition may be administered in vivo to mammals, such as man, to inhibit HCV serine protease or to treat or prevent HCV virus infection.
  • another aspect of this invention provides uses in methods of inhibiting HCV serine protease activity in patients by administering a compound of the present invention or a pharmaceutically acceptable salt or solvate thereof.
  • a compound for use in a method of treating an HCV infection in a patient comprising administering to the patient a therapeutically effective amount of the compound of the invention, or a pharmaceutically acceptable solvate or salt thereof.
  • the method of administering the compound is effective to inhibit the function of the HCV serine protease.
  • the method further comprises administering another compound having anti-HCV activity (as described above) prior to, after or concurrently with a compound of the invention.
  • the compounds of the invention may also be used as laboratory reagents.
  • Compounds may be instrumental in providing research tools for designing of viral replication assays, validation of animal assay systems and structural biology studies to further enhance knowledge of the HCV disease mechanisms. Further, the compounds of the present invention are useful in establishing or determining the binding site of other antiviral compounds, for example, by competitive inhibition.
  • the compounds of this invention may also be used to treat or prevent viral contamination of materials and therefore reduce the risk of viral infection of laboratory or medical personnel or patients who come in contact with such materials, e.g., blood, tissue, surgical instruments and garments, laboratory instruments and garments, and blood collection or transfusion apparatuses and materials.
  • materials e.g., blood, tissue, surgical instruments and garments, laboratory instruments and garments, and blood collection or transfusion apparatuses and materials.
  • the compounds and compositions of the invention can be used for the manufacture of a medicament for treating an HCV infection in a patient.
  • Solution percentages express a weight to volume relationship, and solution ratios express a volume to volume relationship, unless stated otherwise.
  • Nuclear magnetic resonance (NMR) spectra were recorded either on a Bruker 300, 400 or 500 MHz spectrometer; the chemical shifts ( ⁇ ) are reported in parts per million. Flash chromatography was carried out on silica gel (SiO 2 ) according to Still's flash chromatography technique ( W.C. Still et al., J. Org. Chem., (1978), 43, 2923 ).
  • each compound was analyzed by LC/MS, using one of seven methodologies, having the following conditions.
  • the reaction was stirred for 24 h at rt.
  • the dark reaction mixture was partitioned between ether (200 mL) and water (600 mL).
  • the aqueous phase was acidified to pH 4 using 4N HCl.
  • the resulting milky yellow solution was extracted with ether (4 x 200 mL).
  • the combined ether extracts were dried (MgSO 4 ), filtered, and concentrated in vacuo to give a golden oil.
  • the named compound was made racemic by each of the following methods A and B. This racemate could also be resolved to afford chiral Boc-(1 R ,2 S )-1-amino-2-vinylcyclopropyl carboxylic acid ester which was deprotected under acid conditions to afford (1 R , 2 S )-1-amino-2-vinylcyclopropane carboxylic acid ester hydrochloride (Method C).
  • Glycine ethyl ester hydrochloride (303.8 g, 2.16 mole) was suspended in tert- butylmethyl ether (1.6 L). Benzaldehyde (231 g, 2.16 mole) and anhydrous sodium sulfate (154.6 g, 1.09 mole) were added and the mixture cooled to 0 °C using an ice-water bath. Triethylamine (455 mL, 3.26 mole) was added dropwise over 30 min and the mixture stirred for 48 h at rt. The reaction was then quenched by addition of ice-cold water (1 L) and the organic layer was separated.
  • N -Boc-(1 R ,2 S )/(1 S ,2 R )-1-amino-2-vinylcyclopropane carboxylic acid ethyl ester (9.39 g, 36.8 mmol) was dissolved in 4 N HCl/dioxane (90ml, 360 mmol) and was stirred for 2 h at rt. The reaction mixture was concentrated to supply (1 R ,2 S )/(1 S , R )-1-amino-2-vinylcyclopropane carboxylic acid ethyl ester hydrochloride in quanitative yield (7 g, 100%).
  • the enantio-excess of the ester was determined to be 97.2%, and the reaction was cooled to room temperature (26°C) and stirred overnight (16h) after which the enantio-excess of the ester was determined to be 100%.
  • the pH of the reaction mixture was then adjusted to 8.5 with 50% NaOH and the resulting mixture was extracted with MTBE (2 x 2 L).
  • the combined MTBE extract was then washed with 5% NaHCO 3 (3 x 100 mL), water (3 x 100 mL), and evaporated in vacuo to give the enantiomerically pure N -Boc-(1 R ,2 S )/1-amino-2-vinylcyclopropane carboxylic acid ethyl ester as light yellow solid (42.55 g; purity: 97% @ 210 nm, containing no acid; 100% enantiomeric excess ("ee").
  • the aqueous layer from the extraction process was then acidified to pH 2 with 50% H 2 SO 4 and extracted with MTBE (2 x 2 L).
  • enantio-excess of the ester was determined to be 44.3% as following: 0.1 mL of the reaction mixture was removed and mixed well with 1 mL ethanol; after centrifugation, 10 microliter (" ⁇ l") of the supernatant was analyzed with the chiral HPLC. To the remaining reaction mixture, 0.1 mL of DMSO was added, and the plate was incubated for additional 3 days at 250 rpm at 40°C, after which four mL of ethanol was added to the well. After centrifugation, 10 ⁇ l of the supernatant was analyzed with the chiral HPLC and enantio-excess of the ester was determined to be 100%.
  • enantio-excess of the ester was determined to be 39.6% as following: 0.1 mL of the reaction mixture was removed and mixed well with 1 mL ethanol; after cenrifugation, 10 ⁇ l of the supernatant was analyzed with the chiral HPLC. To the remaining reaction mixture, 0.1 mL of DMSO was added, and the plate was incubated for additional 3 days at 250 rpm at 40°C, after which four mL of ethanol was added to the well. After centrifugation, 10 ⁇ l of the supernatant was analyzed with the chiral HPLC and enantio-excess of the ester was determined to be 100%.
  • Step 1 Preparation of 2-Ethylcyclopropane-1,1-dicarboxylic acid di-tert-butyl ester; shown below.
  • Step 2 Preparation of racemic 2-Ethylcyclopropane-1,1-dicarboxylic acid tert-butyl ester, shown below.
  • Step 1 The product of Step 1 (18.3 g, 67.8 mmol) was added to a suspension of potassium tert-butoxide (33.55 g, 299.0 mmol) in dry ether (500 mL) at 0 °C, followed by H 2 O (1.35 mL, 75.0 mmol) and was vigorously stirred overnight at rt.
  • the reaction mixture was poured in a mixture of ice and water and washed with ether (3x).
  • the aqueous layer was acidified with a 10% aq. citric acid solution at 0°C and extracted with EtOAc (3x).
  • the combined organic layers were washed with water (2x), brine, dried (MgSO 4 and concentrated in vacuo to afford the titled product as a pale yellow oil (10 g, 46.8 mmol, 69% yield).
  • Step 3 Preparation of (1 R ,2 R )/(1 S ,2 S ) 2-Ethyl-1-(2-trimethylsilanylethoxycarbonylamino)cyclopropane-carboxylic acid tert-butyl ester, shown below.
  • reaction mixture was filtered, diluted with Et 2 O, washed with a 10% aqueous citric acid solution, water, saturated aqueous NaHCO 3 , water (2x), brine (2X), dried (MgSO 4 ) and concentrated in vacuo.
  • Step 4 Preparation of racemic (1 R ,2 R )/(1 S ,2 S ) 1-Amino-2-ethylcyclopropanecarboxylic acid tert -butyl ester, shown below.
  • Step 3 To the product of Step 3 (3 g, 9 mmol) was added a 1.0 M TBAF solution in THF (9.3 mL, 9.3 mmol) and the mixture heated to reflux for 1.5 h, cooled to rt and then diluted with 500 mL of EtOAc. The solution was successively washed with water (2x100 mL), brine (2x100 mL), dried (MgSO 4 ), concentrated in vacuo to provide the title intermediate
  • the intermediate sulfamoyl chloride was prepared by addition of water (1 equiv) in THF to a cold (-20 °C) stirred solution of chlorosulfonyl isocyanate (1 equiv) in THF and the resulting solution allowed to warm to 0 °C. To this solution was added anhydrous Et 3 N (1 equiv) followed by requisite secondary amine (1 equiv). The reaction mixture was warmed to room temperature, then filtered and the filtrate was rotary evaporated to afford the desired sulfamides.
  • the key step in the preparation of Compounds 1-13 is the coupling of a tripeptide carboxylic acid and a sulfamide to provide the desired tripeptide sulfamide:
  • the general reaction scheme of this key step is shown below:
  • Compound 2 was prepared by the route shown in Scheme 1 of Example 5 and as described for Compound 1 of Example 6. In the case of Compound 2 however, the acylsulfamide employed was synthesized using methoxymethylamine in place of methylethylamine. LC-MS (retention time: 2.070 min.), MS m/z 767 (MH + ).
  • Compound 5 was prepared by the route shown in Scheme 1 of Example 5 and as described for Compound 1 of Example 6. In the case of Compound 5 however, the acylsulfamide employed was synthesized using N-methylaniline in place of methylethylamine. LC-MS (retention time: 2.503 min.), MS m/z 813 (MH + ).
  • Compound 7 was prepared by the route shown in Scheme 1 of Example 5 and as described for Compound 1 of Example 6. In the case of Compound 7 however, the acylsulfamide employed was synthesized using isopropylmethylamie in place of methylethylamine. LC-MS (retention time: 2.420 min.), MS m/z 779 (MH + ).
  • Compound 10 was prepared by the route shown in Scheme 1 of Example 5 and as described for Compound 1 of Example 6. In the case of Compound 10 however, the acylsulfamide employed was synthesized using piperidine in place of methylethylamine. LC-MS (retention time: 2.453 min.), MS m/z 791 (MH + ).
  • Compound 13 was prepared by the route shown in Scheme 1 of Example 5 and as described for Compound 1 of Example 6. In the case of Compound 13 however, the acylsulfamide employed was synthesized using N-methylpiperizine in place of methylethylamine. LC-MS (retention time: 1.923 min.), MS m/z 806 (MH + ).
  • Compound 15 was prepared by the route shown in Scheme 2 of Example 19 and as described for Compound 14 of Example 19. In the case of Compound 15 however, the acylsulfamide employed was synthesized using dimethylamine in place of methylethylamine.. LC-MS (retention time: 2.463 min.), MS m/z 793 (MH + ).
  • Compound 16 was prepared by the route shown in Scheme 2 of Example 19 and as described for Compound 14 of Example 19. In the case of Compound 16 however, the acylsulfamide employed was synthesized using N-methylaniline in place of methylethylamine. LC-MS (retention time: 2.723 min.), MS m/z 856 (MH + ).
  • Compound 17 was prepared by the route shown in Scheme 2 of Example 19 and as described for Compound 14 of Example 19. In the case of Compound 17 however, the acylsulfamide employed was synthesized using dimethylamine in place of methylethylamine, and 6-methoxy-1-chloroisoquinoline was used in place of 2-phenyl-4-chloro-7-methoxyquinoline. LC-MS (retention time: 2.807 min.), MS m/z 717 (MH + ).
  • Compound 18 was prepared by the route shown in Scheme 2 of Example 19 and as described for Compound 14 of Example 19. In the case of Compound 18 however, the acylsulfamide employed was synthesized using pyrrolidine in place of methylethylamine, and 6-methoxy-1-chloroisoquinoline was used in place of 2-phenyl-4-chloro-7-methoxyquinoline. LC-MS (retention time: 2.927 min.), MS m/z 743 (MH + ).
  • Compound 19 was prepared by the route shown in Scheme 2 of Example 19 and as described for Compound 14 of Example 19. In the case of Compound 19 however, 6-methoxy-1-chloroisoquinoline was used in place of 2-phenyl-4-chloro-7-methoxyquinoline.
  • Compound 20 was prepared by the route shown in Scheme 2 of Example 19 and as described for Compound 14 of Example 19.
  • the acylsulfamide employed was synthesized using N-methylaniline in place of methylethylamine, and 6-methoxy-1-chloroisoquinoline was used in place of 2-phenyl-4-chloro-7-methoxyquinoline.
  • Compound 21 was prepared by the route shown in Scheme 2 of Example 19 and as described for Compound 14 of Example 19.
  • the acylsulfamide, employed was synthesized using methylisopropylamine in place of methylethylamine, and 6-methoxy-1-chloroisoquinoline was used in place of 2-phenyl-4-chloro-7-methoxyquinoline.
  • LC-MS retention time: 2.963 min.
  • MS m/z 768 M+Na + ).
  • Compound 22 was prepared by the route shown in Scheme 2 of Example 19 and as described for Compound 14 of Example 19.
  • the acylsulfamide employed was synthesized using methylisopropylamine in place of methylethylamine, and 1-chloroisoquinoline was used in place of 2-phenyl-4-chloro-7-methoxyquinolin LC-MS (retention time: 2.993 min.), MS m/z 737 (M+Na + ).
  • Compound 23 can be prepared by synthesis methods such as described herein.
  • This in vitro assay was to measure the inhibition of HCV NS3 protease complexes, derived from the BMS strain, H77 strain or J416S strain, as described below, by compounds of the present invention. This assay provides an indication of how effective compounds of the present invention would be in inhibiting HCV proteolytic activity
  • Serum from an HCV-infected patient was obtained from Dr. T. Wright, San Francisco Hospital.
  • An engineered full-length cDNA (compliment deoxyribonucleic acid) template of the HCV genome was constructed from DNA fragments obtained by reverse transcription-PCR (RT-PCR) of serum RNA (ribonucleic acid) and using primers selected on the basis of homology between other genotype 1a strains. From the determination of the entire genome sequence, a genotype 1a was assigned to the HCV isolate according to the classification of Simmonds et al. (See P Simmonds, KA Rose, S Graham, SW Chan, F McOmish, BC Dow, EA Follett, PL Yap and H Marsden, J. Clin.
  • the amino acid sequence of the nonstructural region, NS2-5B was shown to be >97% identical to HCV genotype 1a (H77) and 87% identical to genotype 1b (J4L6S).
  • the infectious clones, H77 (1a genotype) and J4L6S (1b genotype) were obtained from R. Purcell (NIH) and the sequences are published in Genbank (AAB67036, see Yanagi,M., Purcell, R.H., Emerson,S.U. and Bukh,J. Proc. Natl. Acad. Sci. U.S.A.
  • the H77 and J4L6S strains were used for production of recombinant NS3/4A protease complexes.
  • DNA encoding the recombinant HCV NS3/4A protease complex (amino acids 1027 to 1711) for these strains were manipulated as described by P. Gallinari et al. (see Gallinari P, Paolini C, Brennan D, Nardi C, Steinkuhler C, De Francesco R. Biochemistry. 38(17):5620-32, (1999 )). Briefly, a three-lysine solubilizing tail was added at the 3'-end of the NS4A coding region.
  • the cysteine in the P1 position of the NS4A-NS4B cleavage site (amino acid 1711) was changed to a glycine to avoid the proteolytic cleavage of the lysine tag. Furthermore, a cysteine to serine mutation was introduced by PCR at amino acid position 1454 to prevent the autolytic cleavage in the NS3 helicase domain.
  • the variant DNA fragment was cloned in the pET21b bacterial expression vector (Novagen) and the US3/4A complex was expressed in Escherichia. coli strain BL21 (DE3) (Invitrogen) following the protocol described by P. Gallinari et al.
  • the cells were resuspended in lysis buffer (10 mL/g) consisting of 25 mM N-(2-Hydroxyethyl)Piperazine-N'-(2-Ethane Sulfonic acid) (HEPES), pH 7.5, 20% glycerol, 500 mM Sodium Chloride (NaCl), 0.5% Triton X-100, 1 microgram/milliliter (" ⁇ g/mL”) lysozyme, 5 mM Magnesium Chloride (MgCl 2 ), 1 ⁇ g/ml DnaseI, 5mM ⁇ -Mercaptoethanol ( ⁇ ME), Protease inhibitor - Ethylenediamine Tetraacetic acid (EDTA) free (Roche), homogenized and incubated for 20 minutes (min) at 4°C.
  • lysis buffer 10 mL/g
  • HEPES N-(2-Hydroxyethyl)Piperazine-N
  • the homogenate was sonicated and clarified by ultra-centrifugation at 235000 g for 1 h at 4°C. Imidazole was added to the supernatant to a final concentration of 15 mM and the pH adjusted to 8.0.
  • the crude protein extract was loaded on a Nickel - Nitrilotriacetic acid (Ni-NTA) column pre-equilibrated with buffer B (25 mM HEPES, pH 8.0, 20% glycerol, 500 mM NaCl, 0.5% Triton X-100, 15 mM imidazole, 5 mM ⁇ ME). The sample was loaded at a flow rate of 1 mL/min.
  • the column was washed with 15 column volumes of buffer C (same as buffer B except with 0.2% Triton X-100).
  • the protein was eluted with 5 column volumes of buffer D (same as buffer C except with 200 mM Imidazole).
  • NS3/4A protease complex-containing fractions were pooled and loaded on a desalting column Superdex-S200 pre-equilibrated with buffer D (25mM HEPES, pH 7.5, 20% glycerol, 300 mM NaCl, 0.2% Triton X-100, 10 mM ⁇ ME). Sample was loaded at a flow rate of 1 mL/min. NS3/4A protease complex-containing fractions were pooled and concentrated to approximately 0.5 mg/ml. The purity of the NS3/4A protease complexes, derived from the BMS, H77 and J4L6S strains, were judged to be greater than 90% by SDS-PAGE and mass spectrometry analyses.
  • the enzyme was stored at -80°C, thawed on ice and diluted prior to use in assay buffer.
  • the substrate used for the NS3/4A protease assay was RET S 1 (Resonance Energy Transfer Depsipeptide Substrate; AnaSpec, Inc. cat # 22991)(FRET peptide), described by Taliani et al. in Anal. Biochem. 240(2):60-67 (1996 ).
  • the sequence of this peptide is loosely based on the NS4A/NS4B natural cleavage site for the HCV NS3 protease except there is an ester linkage rather than an amide bond at the cleavage site.
  • the peptide substrate was incubated with one of the three recombinant NS3/4A protease complexes, in the absence or presence of a compound of the present invention, and the formation of fluorescent reaction product was followed in real time using a Cytofluor Series 4000.
  • HEPES and Glycerol were obtained from GIBCO-BRL.
  • Dimethyl Sulfoxide (DMSO) was obtained from Sigma.
  • ⁇ -Mercaptoethanol was obtained from Bio Rad.
  • Assay buffer 50 mM HEPES, pH 7.5; 0.15 M NaCl; 0.1% Triton; 15% Glycerol;10 mM ⁇ ME.
  • Substrate 2 ⁇ M final concentration (from a 2 mM stock solution in DMSO stored at -20°C).
  • HCV NS3/4A protease type 1a (1b) 2-3 nM final concentration (from a 5 ⁇ M stock solution in 25 mM HEPES, pH 7.5, 20% glycerol, 300 mM NaCl, 0.2% Triton-X100, 10 mM ⁇ ME).
  • the assay was made more sensitive by adding 50 ⁇ g/ml Bovine Serum Albumin (Sigma) to the assay buffer and reducing the end protease concentration to 300 pM.
  • the assay was performed in a 96-well polystyrene black plate from Falcon. Each well contained 25 ⁇ l NS3/4A protease complex in assay buffer, 50 ⁇ l of a compound of the present invention in 10% DMSO/assay buffer and 25 ⁇ l substrate in assay buffer. A control (no compound) was also prepared on the same assay plate. The enzyme complex was mixed with compound or control solution for 1 min before initiating the enzymatic reaction by the addition of substrate. The assay plate was read immediately using the Cytofluor Series 4000 (Perspective Biosystems). The instrument was set to read an emission of 340 nm and excitation of 490 nm at 25°C. Reactions were generally followed for approximately 15 min.
  • the percent inhibition was calculated with the following equation: 100 - ⁇ F inh / ⁇ F con ⁇ x ⁇ 100 where ⁇ F is the change in fluorescence over the linear range of the curve.
  • ⁇ F is the change in fluorescence over the linear range of the curve.
  • IC 50 50% effective concentration
  • the specificity assays were performed to demonstrate the in vitro selectivity of the compounds of the present invention in inhibiting HCV NS3/4A protease complex as compared to other serine or cysteine proteases.
  • the specificities of compounds of the present invention were determined against a variety of serine proteases: human neutrophil elastase (HNE), porcine pancreatic elastase (PPE) and human pancreatic chymotrypsin and one cysteine protease: human liver cathepsin B.
  • HNE human neutrophil elastase
  • PPE porcine pancreatic elastase
  • chymotrypsin human liver cathepsin B.
  • pNA colorimetric p-nitroaniline
  • Each assay included a 2 h enzyme-inhibitor pre-incubation at room temperature followed by addition of substrate and hydrolysis to ⁇ 30% conversion as measured on a Spectramax Pro microplate reader. Compound concentrations varied from 100 to 0.4 ⁇ M depending on their potency.
  • the percentage of inhibition was calculated using the formula: 1 - UV inh - UV blank / UV ctl - UV blank x 100
  • HCV replicon whole cell system was established as described by Lohmann V, Korner F, Koch J, Herian U, Theilmann L, Bartenschlager R., Science 285(5424):110-3 (1999 ). This system enabled us to evaluate the effects of our HCV Protease compounds on HCV RNA replication. Briefly, using the HCV strain 1b sequence described in the Lohmann paper (Assession number:AJ238799), an HCV cDNA was generated encoding the 5' internal ribosome entry site (IRES), the neomycin resistance gene, the EMCV (encephalomyocarditis viurs)-IRES and the HCV nonstructural proteins, NS3-NS5B, and 3' non-translated region (NTR).
  • IRS internal ribosome entry site
  • EMCV encephalomyocarditis viurs
  • Huh7 cells constitutively expressing the HCV replicon, were grown in Dulbecco's Modified Eagle Media (DMEM) containing 10% Fetal calf serum (FCS) and 1 mg/ml G418 (Gibco-BRL). Cells were seeded the night before (1.5 x 10 4 cells/well) in 96-well tissue-culture sterile plates. Compound and no compound controls were prepared in DMEM containing 4% FCS, 1:100 Penicillin / Streptomysin, 1:100 L-glutamine and 5% DMSO in the dilution plate (0.5% DMSO final concentration in the assay). Compound / DMSO mixes were added to the cells and incubated for 4 days at 37°C.
  • DMEM Dulbecco's Modified Eagle Media
  • FCS Fetal calf serum
  • G418 Gibco-BRL
  • lysis assay reagent containing the FRET peptide (RET S1, as described for the in vitro enzyme assay).
  • the lysis assay reagent was made from 5X cell Luciferase cell culture lysis reagent (Promega #E153A) diluted to 1X with distilled water, NaCl added to 150 mM final, the FRET peptide diluted to 10 ⁇ M final from a 2 mM stock in 100% DMSO.
  • the plate was then placed into the Cytofluor 4000 instrument which had been set to 340 nm excitation / 490 nm emission, automatic mode for 21 cycles and the plate read in a kinetic mode. EC 50 determinations were carried out as described for the IC 50 determinations.
  • RNA assays Two different secondary assays were used to confirm EC 50 determinations from the replicon FRET assay. These included a quantitative RNA assay and a transient luciferase cell reporter assay. For the quantitative RNA assay, compound / no compound controls were incubated with the cells as described for the replicon FRET assay. After 4 days, cells were lyzed using the Rneasy kit (Qiagen). Purified total RNA was normalized using RiboGreen ( Jones LJ, Yue ST, Cheung CY, Singer VL, Anal.
  • RNA made to a volume of 5 ⁇ l ( ⁇ 1 ng) was added to a 20 ⁇ l Ready-Mix containing the following: 1.25X Thermoscript reaction mix (containing Magnesium Sulfate and 2-deoxynucleoside 5'-triphosphates (dNTPs)), 3 mM dNTPs, 200 nM forward primer (sequence: 5'-gggagagccatagtggtctgc-3'), 600 nM reverse primer (5'-cccaaatctccaggcattga-3'), 100 nM probe (5'-6-FAM-cggaattgccaggacgaccgg-BHQ-1-3')(FAM: Fluorescein-aminohexyl amidite; BHQ: Black Hole Quencher), 1 ⁇ M Rox reference dye (Invitrogen cat # 12223-012) and Thermoscript Plus Platinum Taq polymerase mixture.
  • Thermoscript reaction mix containing Magnesium Sul
  • the luciferase reporter assay was also used to confirm compound potency in the replicon. Utilization of a replicon luciferase reporter assay was first described by Krieger et al ( Krieger N, Lohmann V, and Bartenschlager R, J. Virol. 75(10):4614-4624 (2001 )). The replicon construct described for our FRET assay was modified by replacing the resistance gene neomycin with the Blasticidin-resistance gene fused to the N-terminus of the humanized form of Renilla luciferase (restriction sites Ascl / Pme 1 used for the subcloning).
  • the adaptive mutation at position 1179 was also introduced ( Blight KJ, Kolykhalov, AA, Rice, CM, Science 290(5498):1972-1974 ).
  • the luciferase reporter assay was set up by seeding huh7 cells the night before at a density of 2 x 10 6 cells per T75 flask. Cells were washed the next day with 7.5 ml Opti-MEM. Following the Invitrogen protocol, 40 ⁇ l DMRIE-C was vortexed with 5 ml Opti-MEM before adding 5 ⁇ g HCV reporter replicon RNA. The mix was added to the washed huh7 cells and left for 4 h at 37°C.
  • % control average luciferase signal in experimental wells + compound average luciferase signal in DMSO control wells - compound
  • Representative compounds of the invention were assessed in the HCV replicon cell assay described above and/or in several of the outlined specificity assays described above.
  • Compound 9 was found to have an IC 50 of 128 nM against the BMS strain of the NS3/4A protease complex in the enzyme assay described above. Similar potency values were obtained with the published H77 (IC 50 of 62 nM) and J4L6S (IC 50 of 41 nM) strains.
  • the EC 50 value in the replicon assay was 403 nM.
  • the compounds inhibited below were tested for biological activity according to the HCV replican cell assay described above and were found to have activities set forth in Table 2 below.
  • the activity ranges were classified into the following groups: for IC 50 , and EC 50 , A (least active) >1.5 ⁇ M; B 0.15-1.5 ⁇ M; C (most active) ⁇ 0.15 ⁇ M.
  • the IC 50 values are from about 0.001-1 ⁇ M, and most preferably less than 0.1 ⁇ M.
  • the EC 50 values are from about 0.001 to 25 ⁇ M, more preferably from about 0.001-1 ⁇ M and most preferably less than about 0.1 ⁇ M.

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Families Citing this family (139)

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Publication number Priority date Publication date Assignee Title
NZ335276A (en) 1996-10-18 2000-09-29 Vertex Pharma Inhibitors of serine proteases, particularly hepatitis C virus (HCV) NS3 (Non Structural Protein 3) protease
SV2003000617A (es) 2000-08-31 2003-01-13 Lilly Co Eli Inhibidores de la proteasa peptidomimetica ref. x-14912m
MY140680A (en) 2002-05-20 2010-01-15 Bristol Myers Squibb Co Hepatitis c virus inhibitors
CN103204903A (zh) * 2003-05-21 2013-07-17 贝林格尔.英格海姆国际有限公司 丙型肝炎抑制剂化合物
MY148123A (en) 2003-09-05 2013-02-28 Vertex Pharma Inhibitors of serine proteases, particularly hcv ns3-ns4a protease
US7491794B2 (en) * 2003-10-14 2009-02-17 Intermune, Inc. Macrocyclic compounds as inhibitors of viral replication
US7132504B2 (en) 2003-11-12 2006-11-07 Bristol-Myers Squibb Company Hepatitis C virus inhibitors
US7135462B2 (en) 2003-11-20 2006-11-14 Bristol-Myers Squibb Company Hepatitis C virus inhibitors
US7309708B2 (en) 2003-11-20 2007-12-18 Birstol-Myers Squibb Company Hepatitis C virus inhibitors
EP1730167B1 (en) * 2004-01-21 2011-01-12 Boehringer Ingelheim International GmbH Macrocyclic peptides active against the hepatitis c virus
EP1763531A4 (en) * 2004-06-28 2009-07-01 Boehringer Ingelheim Int ANALOGUE OF HEPATITIS-C INHIBITING PEPTIDES
JP4914355B2 (ja) * 2004-07-20 2012-04-11 ベーリンガー インゲルハイム インターナショナル ゲゼルシャフト ミット ベシュレンクテル ハフツング Ｃ型肝炎インヒビターペプチド類似体
UY29016A1 (es) * 2004-07-20 2006-02-24 Boehringer Ingelheim Int Analogos de dipeptidos inhibidores de la hepatitis c
EP2374464A3 (en) 2004-10-01 2011-10-26 Vertex Pharmaceuticals Incorporated HCV N3S-NS4A protease inhibition
TW201424733A (zh) 2004-10-29 2014-07-01 Vertex Pharma 劑量型式
US7323447B2 (en) 2005-02-08 2008-01-29 Bristol-Myers Squibb Company Hepatitis C virus inhibitors
WO2006119061A2 (en) 2005-05-02 2006-11-09 Merck & Co., Inc. Hcv ns3 protease inhibitors
US7592336B2 (en) 2005-05-10 2009-09-22 Bristol-Myers Squibb Company Hepatitis C virus inhibitors
CN104436190A (zh) 2005-06-08 2015-03-25 达纳-法伯癌症研究院公司 通过抑制程序性细胞死亡1(pd-1)途经治疗持续性感染和癌症的方法及组合物
US7601686B2 (en) 2005-07-11 2009-10-13 Bristol-Myers Squibb Company Hepatitis C virus inhibitors
TWI389908B (zh) * 2005-07-14 2013-03-21 Gilead Sciences Inc 抗病毒化合物類
US20080200497A1 (en) * 2005-07-20 2008-08-21 Bailey Murray D Hepatitis C Inhibitor Peptide Analogs
NZ594105A (en) 2005-07-25 2013-02-22 Intermune Inc Novel macrocyclic inhibitors of hepatitis c virus replication
CA2615896C (en) 2005-08-01 2012-11-13 Merck & Co., Inc. Macrocyclic peptides as hcv ns3 protease inhibitors
WO2007019674A1 (en) 2005-08-12 2007-02-22 Boehringer Ingelheim International Gmbh Viral polymerase inhibitors
US7964624B1 (en) 2005-08-26 2011-06-21 Vertex Pharmaceuticals Incorporated Inhibitors of serine proteases
AR055395A1 (es) 2005-08-26 2007-08-22 Vertex Pharma Compuestos inhibidores de la actividad de la serina proteasa ns3-ns4a del virus de la hepatitis c
ATE493409T1 (de) 2005-10-11 2011-01-15 Intermune Inc Verbindungen und verfahren zur inhibierung der replikation des hepatitis-c-virus
US7772183B2 (en) 2005-10-12 2010-08-10 Bristol-Myers Squibb Company Hepatitis C virus inhibitors
US7741281B2 (en) 2005-11-03 2010-06-22 Bristol-Myers Squibb Company Hepatitis C virus inhibitors
US7816348B2 (en) 2006-02-03 2010-10-19 Boehringer Ingelheim International Gmbh Viral polymerase inhibitors
CN102614490A (zh) 2006-02-27 2012-08-01 弗特克斯药品有限公司 包含vx-950的共晶体和包含所述共晶体的药物组合物
EP2194039A1 (en) 2006-03-16 2010-06-09 Vertex Pharmceuticals Incorporated Process for preparing optically enriched compounds
GB0609492D0 (en) 2006-05-15 2006-06-21 Angeletti P Ist Richerche Bio Therapeutic agents
US20080187516A1 (en) * 2006-06-06 2008-08-07 Ying Sun Acyclic oximyl hepatitis c protease inhibitors
US7728148B2 (en) * 2006-06-06 2010-06-01 Enanta Pharmaceuticals, Inc. Acyclic oximyl hepatitis C protease inhibitors
GB0612423D0 (en) 2006-06-23 2006-08-02 Angeletti P Ist Richerche Bio Therapeutic agents
AR061629A1 (es) * 2006-06-26 2008-09-10 Enanta Pharm Inc Quinoxalinil macrociclicos inhibidores de serina proteasa del virus de la hepatitis c. proceso de obtencion y composiciones farmaceuticas
KR20090024834A (ko) 2006-07-05 2009-03-09 인터뮨, 인크. Ｃ형 간염 바이러스 복제의 신규 억제제
EP2049474B1 (en) 2006-07-11 2015-11-04 Bristol-Myers Squibb Company Hepatitis c virus inhibitors
EP2374812B1 (en) 2006-07-13 2015-04-15 Achillion Pharmaceuticals, Inc. 4-amino-4-oxobutanoyl Peptides as Inhibitors of Viral Replication
US20090035267A1 (en) * 2007-07-31 2009-02-05 Moore Joel D Acyclic, pyridazinone-derived hepatitis c serine protease inhibitors
US7635683B2 (en) * 2006-08-04 2009-12-22 Enanta Pharmaceuticals, Inc. Quinoxalinyl tripeptide hepatitis C virus inhibitors
US7687459B2 (en) * 2006-08-11 2010-03-30 Enanta Pharmaceuticals, Inc. Arylalkoxyl hepatitis C virus protease inhibitors
US20080038225A1 (en) * 2006-08-11 2008-02-14 Ying Sun Triazolyl acyclic hepatitis c serine protease inhibitors
EP1886685A1 (en) 2006-08-11 2008-02-13 INSERM (Institut National de la Santé et de la Recherche Médicale) Methods, uses and compositions for modulating replication of hcv through the farnesoid x receptor (fxr) activation or inhibition
EP2054388A4 (en) 2006-08-17 2009-10-28 Boehringer Ingelheim Int VIRAL POLYMERASE HEMMER
CA2667266C (en) 2006-10-24 2015-11-24 Merck & Co., Inc. Hcv ns3 protease inhibitors
JP2010507656A (ja) 2006-10-24 2010-03-11 メルク エンド カムパニー インコーポレーテッド Ｈｃｖｎｓ３プロテアーゼ阻害剤
US8138164B2 (en) 2006-10-24 2012-03-20 Merck Sharp & Dohme Corp. HCV NS3 protease inhibitors
US8377874B2 (en) 2006-10-27 2013-02-19 Merck Sharp & Dohme Corp. HCV NS3 protease inhibitors
JP5268927B2 (ja) 2006-10-27 2013-08-21 メルク・シャープ・アンド・ドーム・コーポレーション Ｈｃｖｎｓ３プロテアーゼ阻害剤
US8343477B2 (en) 2006-11-01 2013-01-01 Bristol-Myers Squibb Company Inhibitors of hepatitis C virus
US7772180B2 (en) 2006-11-09 2010-08-10 Bristol-Myers Squibb Company Hepatitis C virus inhibitors
US7763584B2 (en) 2006-11-16 2010-07-27 Bristol-Myers Squibb Company Hepatitis C virus inhibitors
US8003604B2 (en) 2006-11-16 2011-08-23 Bristol-Myers Squibb Company Hepatitis C virus inhibitors
US7888464B2 (en) 2006-11-16 2011-02-15 Bristol-Myers Squibb Company Hepatitis C virus inhibitors
GB0625349D0 (en) 2006-12-20 2007-01-31 Angeletti P Ist Richerche Bio Therapeutic compounds
AU2007335962B2 (en) 2006-12-20 2012-09-06 Istituto Di Ricerche Di Biologia Molecolare P. Angeletti Spa Antiviral indoles
GB0625345D0 (en) 2006-12-20 2007-01-31 Angeletti P Ist Richerche Bio Therapeutic compounds
EP2125113A2 (en) 2007-02-26 2009-12-02 Achillion Pharmaceuticals, Inc. Tertiary amine substituted peptides useful as inhibitors of hcv replication
EP2134717A2 (en) 2007-02-27 2009-12-23 Vertex Pharmceuticals Incorporated Inhibitors of serine proteases
WO2008106151A2 (en) 2007-02-27 2008-09-04 Vertex Pharmaceuticals Incorporated Co-crystals and pharmaceutical compositions comprising the same
PT2144604E (pt) 2007-02-28 2011-10-19 Conatus Pharmaceuticals Inc Métodos para o tratamento da hepatite c viral crónica utilizando ro-113-0830
US20080267917A1 (en) * 2007-04-26 2008-10-30 Deqiang Niu N-functionalized amides as hepatitis c serine protease inhibitors
CA2686546A1 (en) * 2007-05-10 2008-11-20 Intermune, Inc. Novel peptide inhibitors of hepatitis c virus replication
US20090005387A1 (en) * 2007-06-26 2009-01-01 Deqiang Niu Quinoxalinyl macrocyclic hepatitis c virus serine protease inhibitors
AU2008271116B2 (en) 2007-06-29 2012-09-20 Gilead Sciences, Inc. Antiviral compounds
CN101801925A (zh) 2007-06-29 2010-08-11 吉里德科学公司 抗病毒组合物
EP2178885A1 (en) 2007-07-17 2010-04-28 Istituto Di Ricerche Di Biologia Molecolare P. Angeletti S.P.A. Macrocyclic indole derivatives for the treatment of hepatitis c infections
US8927569B2 (en) 2007-07-19 2015-01-06 Merck Sharp & Dohme Corp. Macrocyclic compounds as antiviral agents
CA2693997C (en) 2007-08-03 2013-01-15 Pierre L. Beaulieu Viral polymerase inhibitors
CL2008002549A1 (es) * 2007-08-30 2010-09-03 Vertex Pharma Cocristal que comprende vx-950 y un formador de cocristal seleccionado de acido 3-metoxi-4hidroxibenzoico,acido 2,4-dihidroxibenzoico y acido 2,5-dihidroxibenzoico; metodo de preparacion; composicion farmaceutica que comprende el cocristal, util como agente antiviral en el tratamiento del hcv.
CA2700383A1 (en) 2007-09-24 2009-04-02 Achillion Pharmaceuticals, Inc. Urea-containing peptides as inhibitors of viral replication
US8030307B2 (en) * 2007-11-29 2011-10-04 Enanta Pharmaceuticals, Inc. Bicyclic, C5-substituted proline derivatives as inhibitors of the hepatitis C virus NS3 protease
WO2009070692A1 (en) * 2007-11-29 2009-06-04 Enanta Pharmaceuticals, Inc. C5-substituted, proline-derived, macrocyclic hepatitis c serine protease inhibitors
US8361958B2 (en) * 2007-12-05 2013-01-29 Enanta Pharmaceuticals, Inc. Oximyl HCV serine protease inhibitors
WO2009076173A2 (en) 2007-12-05 2009-06-18 Enanta Pharmaceuticals, Inc. Fluorinated tripeptide hcv serine protease inhibitors
CN101903351B (zh) 2007-12-19 2014-09-10 贝林格尔.英格海姆国际有限公司 病毒聚合酶抑制剂
EP2237666A4 (en) * 2007-12-21 2012-05-16 Avila Therapeutics Inc HCV PROTEASE INHIBITORS AND USES THEREOF
WO2009082697A1 (en) * 2007-12-21 2009-07-02 Avila Therapeutics, Inc. Hcv protease inhibitors and uses thereof
US8202996B2 (en) 2007-12-21 2012-06-19 Bristol-Myers Squibb Company Crystalline forms of N-(tert-butoxycarbonyl)-3-methyl-L-valyl-(4R)-4-((7-chloro-4-methoxy-1-isoquinolinyl)oxy)-N- ((1R,2S)-1-((cyclopropylsulfonyl)carbamoyl)-2-vinylcyclopropyl)-L-prolinamide
US8309685B2 (en) * 2007-12-21 2012-11-13 Celgene Avilomics Research, Inc. HCV protease inhibitors and uses thereof
US8293705B2 (en) 2007-12-21 2012-10-23 Avila Therapeutics, Inc. HCV protease inhibitors and uses thereof
EP2245015A4 (en) * 2008-01-24 2012-05-30 Enanta Pharm Inc DIFLUORATED TRIPEPTIDES AS INHIBITORS OF HCV SERINE PROTEASE
EP2252311A1 (en) * 2008-01-24 2010-11-24 Enanta Pharmaceuticals, Inc. Heteroaryl-containing tripeptide hcv serine protease inhibitors
KR100945784B1 (ko) 2008-01-24 2010-03-08 한국외국어대학교 연구산학협력단 Ｃ형 간염 바이러스의 중합효소 ｎｓ５ｂ를 저해하는 c형 간염 바이러스 치료제
MX2010008523A (es) 2008-02-04 2010-08-31 Idenix Pharmaceuticals Inc Inhibidores macrociclicos de serina proteasa.
EP2268619A4 (en) * 2008-03-20 2012-04-04 Enanta Pharm Inc FLUORINATED MACROCYCLIC COMPOUNDS AS INHIBITORS OF HEPATITIS C VIRUS
SG175692A1 (en) 2008-04-15 2011-11-28 Intermune Inc Novel macrocyclic inhibitors of hepatitis c virus replication
US8163921B2 (en) 2008-04-16 2012-04-24 Bristol-Myers Squibb Company Hepatitis C virus inhibitors
AU2009241445A1 (en) 2008-04-28 2009-11-05 Merck Sharp & Dohme Corp. HCV NS3 protease inhibitors
US8211891B2 (en) * 2008-04-30 2012-07-03 Enanta Pharmaceuticals, Inc. Difluoromethyl-containing macrocyclic compounds as hepatitis C virus inhibitors
CN102046648A (zh) 2008-05-29 2011-05-04 百时美施贵宝公司 丙型肝炎病毒抑制剂
US7964560B2 (en) 2008-05-29 2011-06-21 Bristol-Myers Squibb Company Hepatitis C virus inhibitors
DK2310095T3 (da) 2008-07-22 2012-12-10 Merck Sharp & Dohme Makrocykliske quinoxalinforbindelser som hcv-ns3-protease-inhibitorer
US8207341B2 (en) 2008-09-04 2012-06-26 Bristol-Myers Squibb Company Process or synthesizing substituted isoquinolines
UY32099A (es) 2008-09-11 2010-04-30 Enanta Pharm Inc Inhibidores macrocíclicos de serina proteasas de hepatitis c
US8044087B2 (en) 2008-09-29 2011-10-25 Bristol-Myers Squibb Company Hepatitis C virus inhibitors
US8563505B2 (en) 2008-09-29 2013-10-22 Bristol-Myers Squibb Company Hepatitis C virus inhibitors
MX2011005691A (es) 2008-11-28 2011-07-20 Univ Emory Metodos para el tratamiento de infecciones y tumores.
AU2009324643B2 (en) 2008-12-10 2014-08-28 Achillion Pharmaceuticals, Inc. New 4-amino-4-oxobutanoyl peptides as inhibitors of viral replication
US8283310B2 (en) 2008-12-15 2012-10-09 Bristol-Myers Squibb Company Hepatitis C virus inhibitors
US20100160403A1 (en) 2008-12-19 2010-06-24 Gilead Sciences, Inc. Hcv ns3 protease inhibitors
MX2011007195A (es) 2009-01-07 2013-07-12 Scynexis Inc Derivado de ciclosporina para el uso en el tratamiento de infección de virus de hepatitis c (vhc) y virus de inmunodeficiencia humana (vih).
EP2396028A2 (en) 2009-02-12 2011-12-21 Vertex Pharmceuticals Incorporated Hcv combination therapies comprising pegylated interferon, ribavirin and telaprevir
AR075584A1 (es) 2009-02-27 2011-04-20 Intermune Inc COMPOSICIONES TERAPEUTICAS QUE COMPRENDEN beta-D-2'-DESOXI-2'-FLUORO-2'-C-METILCITIDINA Y UN DERIVADO DE ACIDO ISOINDOL CARBOXILICO Y SUS USOS. COMPUESTO.
US8377962B2 (en) 2009-04-08 2013-02-19 Idenix Pharmaceuticals, Inc. Macrocyclic serine protease inhibitors
JP5639155B2 (ja) 2009-05-13 2014-12-10 エナンタ ファーマシューティカルズ インコーポレイテッド Ｃ型肝炎ウイルスインヒビターとしての大環状化合物
US8232246B2 (en) 2009-06-30 2012-07-31 Abbott Laboratories Anti-viral compounds
WO2011014487A1 (en) 2009-07-30 2011-02-03 Merck Sharp & Dohme Corp. Hepatitis c virus ns3 protease inhibitors
AR077712A1 (es) 2009-08-05 2011-09-14 Idenix Pharmaceuticals Inc Inhibidores de serina proteasa macrociclica
WO2011049908A2 (en) * 2009-10-19 2011-04-28 Enanta Pharmaceuticals, Inc. Bismacrokyclic compounds as hepatitis c virus inhibitors
WO2011094489A1 (en) 2010-01-29 2011-08-04 Vertex Pharmaceuticals Incorporated Therapies for treating hepatitis c virus infection
WO2011156545A1 (en) 2010-06-09 2011-12-15 Vertex Pharmaceuticals Incorporated Viral dynamic model for hcv combination therapy
AR082215A1 (es) 2010-07-14 2012-11-21 Vertex Pharma Composicion farmaceutica agradable al paladar
DK2618831T3 (en) 2010-09-21 2016-04-04 Enanta Pharm Inc Macrocyclic prolinafledte HCV serine protease inhibitors
WO2012048235A1 (en) * 2010-10-08 2012-04-12 Novartis Ag Vitamin e formulations of sulfamide ns3 inhibitors
BR112013016480A2 (pt) 2010-12-30 2016-09-20 Abbvie Inc macrocíclo da fenantridina inibadores da protease da serina da hepatite c
US8937041B2 (en) 2010-12-30 2015-01-20 Abbvie, Inc. Macrocyclic hepatitis C serine protease inhibitors
TW201309690A (zh) 2011-02-10 2013-03-01 Idenix Pharmaceuticals Inc 巨環絲胺酸蛋白酶抑制劑，其醫藥組合物及其於治療ｈｃｖ感染之用途
WO2012107589A1 (en) 2011-02-11 2012-08-16 INSERM (Institut National de la Santé et de la Recherche Médicale) Methods and pharmaceutical compositions for the treatment and prevention of hcv infections
WO2012109646A1 (en) 2011-02-11 2012-08-16 Vertex Pharmaceuticals Incorporated Treatment of hcv in hiv infection patients
US8957203B2 (en) 2011-05-05 2015-02-17 Bristol-Myers Squibb Company Hepatitis C virus inhibitors
US10201584B1 (en) 2011-05-17 2019-02-12 Abbvie Inc. Compositions and methods for treating HCV
US8691757B2 (en) 2011-06-15 2014-04-08 Bristol-Myers Squibb Company Hepatitis C virus inhibitors
EP2780026B1 (en) 2011-11-15 2019-10-23 Merck Sharp & Dohme Corp. Hcv ns3 protease inhibitors
US20130195797A1 (en) 2012-01-31 2013-08-01 Vertex Pharmaceuticals Incorporated High potency formulations of vx-950
JP6154474B2 (ja) 2012-10-19 2017-06-28 ブリストル−マイヤーズ スクイブ カンパニーＢｒｉｓｔｏｌ−Ｍｙｅｒｓ Ｓｑｕｉｂｂ Ｃｏｍｐａｎｙ Ｃ型肝炎ウイルス阻害剤
WO2014070964A1 (en) 2012-11-02 2014-05-08 Bristol-Myers Squibb Company Hepatitis c virus inhibitors
US9643999B2 (en) 2012-11-02 2017-05-09 Bristol-Myers Squibb Company Hepatitis C virus inhibitors
EP2914598B1 (en) 2012-11-02 2017-10-18 Bristol-Myers Squibb Company Hepatitis c virus inhibitors
EP2914614B1 (en) 2012-11-05 2017-08-16 Bristol-Myers Squibb Company Hepatitis c virus inhibitors
EP2964664B1 (en) 2013-03-07 2017-01-11 Bristol-Myers Squibb Company Hepatitis c virus inhibitors
EP2970195B1 (en) 2013-03-14 2017-08-02 Achillion Pharmaceuticals, Inc. Processes for producing sovaprevir
KR20160005686A (ko) 2013-03-15 2016-01-15 아칠리온 파르마세우티칼스 인코포레이티드 소바프레비르 다형체들 및 이의 제조 방법
US9085607B2 (en) 2013-03-15 2015-07-21 Achillion Pharmaceuticals, Inc. ACH-0142684 sodium salt polymorph, composition including the same, and method of manufacture thereof
WO2014145507A1 (en) 2013-03-15 2014-09-18 Achillion Pharmaceuticals, Inc. A process for making a 4-amino-4-oxobutanoyl peptide cyclic analogue, an inhibitor of viral replication, and intermediates thereof
EP3089757A1 (en) 2014-01-03 2016-11-09 AbbVie Inc. Solid antiviral dosage forms

Family Cites Families (43)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5610054A (en) 1992-05-14 1997-03-11 Ribozyme Pharmaceuticals, Inc. Enzymatic RNA molecule targeted against Hepatitis C virus
US5500208A (en) 1994-06-07 1996-03-19 The Procter & Gamble Company Oral compositions comprising a novel tripeptide
MY147327A (en) 1995-06-29 2012-11-30 Novartis Ag Somatostatin peptides
GB9517022D0 (en) 1995-08-19 1995-10-25 Glaxo Group Ltd Medicaments
US5633388A (en) 1996-03-29 1997-05-27 Viropharma Incorporated Compounds, compositions and methods for treatment of hepatitis C
EP0907659A1 (en) 1996-05-10 1999-04-14 Schering Corporation Synthetic inhibitors of hepatitis c virus ns3 protease
NZ335276A (en) 1996-10-18 2000-09-29 Vertex Pharma Inhibitors of serine proteases, particularly hepatitis C virus (HCV) NS3 (Non Structural Protein 3) protease
GB9623908D0 (en) 1996-11-18 1997-01-08 Hoffmann La Roche Amino acid derivatives
AU7127298A (en) 1997-04-14 1998-11-11 Emory University Serine protease inhibitors
GB9707659D0 (en) 1997-04-16 1997-06-04 Peptide Therapeutics Ltd Hepatitis C NS3 Protease inhibitors
IL134232A0 (en) 1997-08-11 2001-04-30 Boehringer Ingelheim Ca Ltd Hepatitis c inhibitor peptides
ES2234144T3 (es) 1997-08-11 2005-06-16 Boehringer Ingelheim (Canada) Ltd. Analogos de peptidos inhibidores de la hepatitis c.
IT1299134B1 (it) 1998-02-02 2000-02-29 Angeletti P Ist Richerche Bio Procedimento per la produzione di peptidi con proprieta' inibitrici della proteasi ns3 del virus hcv, peptidi cosi' ottenibili e peptidi
AU3376699A (en) 1998-03-31 1999-10-18 Vertex Pharmaceuticals Incorporated Inhibitors of serine proteases, particularly hepatitis c virus ns3 protease
GB9812523D0 (en) 1998-06-10 1998-08-05 Angeletti P Ist Richerche Bio Peptide inhibitors of hepatitis c virus ns3 protease
DE69925918T2 (de) 1998-07-27 2006-05-11 Istituto Di Ricerche Di Biologia Molecolare P. Angeletti S.P.A. Diketosäure-derivate als hemmstoffe von polymerasen
AR022061A1 (es) 1998-08-10 2002-09-04 Boehringer Ingelheim Ca Ltd Peptidos inhibidores de la hepatitis c, una composicion farmaceutica que los contiene, el uso de los mismos para preparar una composicion farmaceutica, el uso de un producto intermedio para la preparacion de estos peptidos y un procedimiento para la preparacion de un peptido analogo de los mismos.
US6323180B1 (en) 1998-08-10 2001-11-27 Boehringer Ingelheim (Canada) Ltd Hepatitis C inhibitor tri-peptides
EP1128832A4 (en) 1998-08-21 2003-03-05 Viropharma Inc COMPOUNDS, COMPOSITIONS AND METHODS FOR TREATING VIRAL INFECTIONS AND RELATED DISEASES
AU751201B2 (en) 1998-09-04 2002-08-08 Viropharma Incorporated Methods for treating or preventing viral infections and associated diseases
EP1115286A4 (en) 1998-09-25 2003-07-23 Viropharma Inc METHOD FOR TREATING AND PREVENTING VIRAL INFECTIONS AND DISEASES RELATED TO THEM
UA74546C2 (en) 1999-04-06 2006-01-16 Boehringer Ingelheim Ca Ltd Macrocyclic peptides having activity relative to hepatitis c virus, a pharmaceutical composition and use of the pharmaceutical composition
US6566365B1 (en) 1999-11-04 2003-05-20 Biochem Pharma Inc. Method for the treatment of Flaviviridea viral infection using nucleoside analogues
SK13752001A3 (sk) 1999-12-27 2002-07-02 Japan Tobacco, Inc. Zlúčeniny s fúzovanými kruhmi a ich použitie ako liečiv
AU2001261377A1 (en) 2000-05-10 2001-11-20 Smith Kline Beecham Corporation Novel anti-infectives
US6448281B1 (en) 2000-07-06 2002-09-10 Boehringer Ingelheim (Canada) Ltd. Viral polymerase inhibitors
GB0017676D0 (en) 2000-07-19 2000-09-06 Angeletti P Ist Richerche Bio Inhibitors of viral polymerase
ES2263687T3 (es) 2000-11-20 2006-12-16 Bristol-Myers Squibb Company Inhibidores tripeptidicos de la hepatitis c.
JP4662115B2 (ja) * 2002-01-24 2011-03-30 独立行政法人科学技術振興機構 フローティングゲートｍｏｓｆｅｔを用いた非線形抵抗回路
CA2369711A1 (en) 2002-01-30 2003-07-30 Boehringer Ingelheim (Canada) Ltd. Macrocyclic peptides active against the hepatitis c virus
CA2370396A1 (en) 2002-02-01 2003-08-01 Boehringer Ingelheim (Canada) Ltd. Hepatitis c inhibitor tri-peptides
CA2369970A1 (en) 2002-02-01 2003-08-01 Boehringer Ingelheim (Canada) Ltd. Hepatitis c inhibitor tri-peptides
US6642204B2 (en) 2002-02-01 2003-11-04 Boehringer Ingelheim International Gmbh Hepatitis C inhibitor tri-peptides
US6828301B2 (en) 2002-02-07 2004-12-07 Boehringer Ingelheim International Gmbh Pharmaceutical compositions for hepatitis C viral protease inhibitors
AU2003301959A1 (en) 2002-05-20 2004-06-03 Bristol-Myers Squibb Company Substituted cycloalkyl p1' hepatitis c virus inhibitors
MY140680A (en) 2002-05-20 2010-01-15 Bristol Myers Squibb Co Hepatitis c virus inhibitors
PL211889B1 (pl) 2002-05-20 2012-07-31 Bristol Myers Squibb Co Pochodna heterocyklosulfonamidowa, kompozycja ją zawierająca oraz ich zastosowanie
PL215228B1 (pl) 2002-05-20 2013-11-29 Bristol Myers Squibb Co Zwiazki tripeptydowe, ich zastosowanie i kompozycja farmaceutyczna je zawierajaca oraz jej zastosowanie
WO2004101605A1 (en) * 2003-03-05 2004-11-25 Boehringer Ingelheim International Gmbh Hepatitis c inhibiting compounds
JP4733023B2 (ja) * 2003-04-16 2011-07-27 ブリストル−マイヤーズ スクイブ カンパニー Ｃ型肝炎ウイルスの大環状イソキノリンペプチド阻害剤
CN103204903A (zh) * 2003-05-21 2013-07-17 贝林格尔.英格海姆国际有限公司 丙型肝炎抑制剂化合物
PE20050431A1 (es) * 2003-09-22 2005-07-19 Boehringer Ingelheim Int Peptidos macrociclicos activos contra el virus de la hepatitis c
US7132504B2 (en) 2003-11-12 2006-11-07 Bristol-Myers Squibb Company Hepatitis C virus inhibitors

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